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2024-03-29T08:46:15Z
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http://2013.igem.org/Team:Tokyo-NoKoGen/oscillator
Team:Tokyo-NoKoGen/oscillator
2013-10-16T09:20:54Z
<p>Jinhee: </p>
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<title>Team:Tokyo-NoKoGen - 2013.igem.org</title><br />
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<link rel="stylesheet" type="text/css" href="Tokyo-NoKoGen.css"><br />
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</head><br />
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<body><br />
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<div id="wrapper"><br />
<br />
<div id="title_logo"> <br />
<br />
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<a href="https://2013.igem.org"><img class="iGEM" src="https://static.igem.org/mediawiki/2013/5/58/IGEM.png"></a> <br />
</div><br />
<br />
<div id="mymenubar"><br />
<br />
<table><br />
<tbody><br />
<tr><br />
<td class="home"><a href="https://2013.igem.org/Team:Tokyo-NoKoGen"><span class="home"></span></a></td><br />
<br />
<td class="project1"><div class="project1"></div></td><br />
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<td class="team"><a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Team"><span class="team"></span></a></td><br />
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<td class="biobrick"><a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Parts"><span class="biobrick"></span></a></td><br />
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<td class="notebook"><a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Notebook"><span class="notebook"></span></a></td><br />
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<td class="humanpractice"><a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Humanpractice"><span class="humanpractice"></span></a></td> <br />
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<td class="achievement"><a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Achievement"><span class="achievement"></span></a></td><br />
</tbody><br />
</table> <br />
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</div><br />
<br />
<div id="index"><br />
<br />
<br />
<h1 id="index_title"></h1><br />
<ul id="contents"><br />
<li><a href="#Introduction"><strong>Introduction</strong></a></li><br />
<li><a href="#Objective"><strong>Objective</strong></a></li><br />
<li><a href="#Method"><strong>Method</strong></a><br />
<ul><br />
<li><a href="#Design ">Design </a></li><br />
<li><a href="#Parts construction">Parts construction</a></li><br />
</ul><br />
</li> <br />
<li><a href="#Evaluation"><strong>Evaluation</strong></a><br />
<ul><br />
<li><a href="#TR-HHR">TR-HHR</a></li><br />
<li><a href="#TR-HHR-taRNA">TR-HHR-taRNA</a></li><br />
</ul><br />
</li><br />
<li><a href="#Future work"><strong>Future work</strong></a></li><br />
<br />
</ul><br />
<br />
<ul id="contents2"><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/oscillator"><li>RNA oscillator</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/scaffold"><li>RNAScaffold</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/light"><li>Light sensor</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/modeling"><li>Modeling</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/rhodopsin"><li>Improving a BioBrick part - Rhodopsin</li></a><br />
</ul><br />
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</div><br />
<div id="main"><br />
<br />
<font size=5><br />
<BR><br />
<p align=center><font size=7><strong>RNA oscillator</strong></font><hr></p><br />
<BR><br />
<BR><br />
<BR><br />
<font size=6><strong>Introduction </span></strong></font><br />
<BR><br />
<BR><br />
<BR><br />
<font size=6 ><strong>Protein oscillator</span></strong></font><br />
<BR><br />
<p style="line-height:110%"><br />
<BR><br />
The oscillating network was built by using three transcriptional repressor systems in which the first repressor protein suppresses the expression of second repressor gene, whose protein suppresses the transcription of the third gene, and the protein the third gene code suppresses the first gene expression.<br />
</p><br />
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<BR><br />
<BR><br />
<br />
<font size=6><strong>Hummer headed ribozyme</span></strong></font><br />
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<BR><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
Hummerheaded ribozymes (HHRs) are found in many species and known to have a capability to self-cleave. Recently, engineered HHRs responding to trans-acting RNAs (taRNAs) in <I>E.coli</I> were reported. In absence of taRNA, the HHRs can self-cleave, but in presence of taRNA, their capability of self-cleavage is inhibited due to hybridize with taRNA and change their structures.<br />
</p><br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/3/3a/%E3%82%AD%E3%83%A3%E3%83%97%E3%83%81%E3%83%A318.PNG><BR><br />
<br />
<BR><BR><BR><br />
<br />
<p style="line-height:110%"><br />
We developed the new oscillator system that is composed only RNAs not using repressor gene and protein. We created three taRNA connected hummerheaded ribozymes (HHR-taRNA) that sense to another taRNA. The same way as the protein oscillator, three HHR-taRNAs inhibits other one’s self-cleavage such as three-way standoff. When the first HHR-taRNA is self-cleaved, connected the first taRNA that binds to the second HHR-taRNA is free and suppresses the second HHR-taRNA self-cleavage. When second HHR-taRNA is self-cleaved, the second taRNA that binds to the third HHR-taRNA is free and suppresses the third HHR-taRNA. Finally, when third HHR-taRNA is self-cleaved, the third taRNA that binds to the first HHR-taRNA is free and suppresses the first HHR-taRNA, completing cycle.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
<font size=6 id="Objective"><strong>Objective</span></strong></font><br />
<BR><br />
<BR><br />
Construction of Three of orthogonal taRNA generator which repressed by another taRNA.<br />
<br />
<BR><br />
<br />
<p style="line-height:110%"><br />
taRNA responsive HHR (TR-HHR) was fused with taRNA. By using orthogonal riboregulator sequences, we designed three of taRNA generators.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
<font size=6 id="Method"><strong>Method</span></strong></font><br />
<br />
<BR><BR><br />
<br />
<font size=5 id="Design"><strong>-Design</span></strong></font> <br />
<br />
<BR><BR><br />
<br />
<p style="line-height:110%"><br />
TR-HHR was designed by Benedikt Klauser and Jo¨ rg S. Hartig. Their TR-HHR were inspired by the riboregulatory system previously reported by Collins and coworkers. They made use of sequence domains of the riboregulator RR12. And, orthogonal variants of riboregulator RR12 (RR12y, RR42) were reported by Collins and coworkers. So, by using the sequences of these orthogonal riboregulators, we designed three orthogonal TR-HHRs.<br />
</p><br />
<br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/a/a1/Rrvariants.png" width="400px" / ><br />
<img src="https://static.igem.org/mediawiki/2013/c/ce/Cccc.png" width="200px" / ><br />
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<BR><BR><br />
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<BR><BR><BR><br />
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<font size=5 id="Parts construction"><strong>-Parts construction</span></strong></font> <br />
<BR><br />
<p style="line-height:110%"><br />
1. Construction of TR-HHR<BR><br />
2. Construction of TR-HHR variants<BR><br />
3. Construction of taRNA variants<BR><br />
4. Construction of TR-HHR-taRNA<BR><br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
1. Construction of TR-HHR [BBa_K1053000]<BR><br />
<br />
<p style="line-height:110%"> <br />
We used the TR-HHR developed by Klauser <I>et al.</I>[1] Two overlap primers designed, and PCR was performed to connect them. The PCR product was ligated into pSB1C3 vector. <br />
</p><br />
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<img src="https://static.igem.org/mediawiki/igem.org/e/e3/Wiki1.png" width="400px" / ><br />
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<BR><BR><br />
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2.Construction of TR-HHR variants<br />
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<BR><BR><BR><br />
<br />
<p style="line-height:110%"><br />
By inverse PCR using pSB1C3-TR-HHR as a template, we could obtain three TR-HHRs, TR(12)-HHR, TR(12y)-HHR, and TR(42)-HHR.<br />
</p><br />
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<img src="https://static.igem.org/mediawiki/igem.org/9/98/Wiki6.png" width="400px" / ><br />
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<BR><br />
<BR><br />
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3.Construction of taRNA variants<br />
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<BR><br />
<BR><br />
<BR><br />
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<p style="line-height:110%"><br />
By inverse PCR using pSB1A3-taR12-DT as a template, we could obtain three taRNAs, taR12, taR12y, taR42.<br />
</p><br />
<br />
<img src="https://static.igem.org/mediawiki/igem.org/a/a7/Wiki2.png" width="400px" / ><br />
<br />
<BR><br />
<BR><br />
<BR><br />
<br />
4.Construction of TR-HHR-taRNA<br />
<BR><BR><br />
[BBa_K1053006][BBa_K1053015][BBa_K1053016]<br />
<BR><br />
<BR><br />
<br />
TR-HHR-taRNAs were constructed by overlap PCR.<br />
<br />
<BR><br />
<BR><br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/9/91/Wiki3.png" width="400px" / ><br />
<br />
<BR><br />
<BR><br />
<BR><br />
<BR><br />
<BR><br />
<BR><br />
<font size=6 id="Evaluation"><strong>Evaluation</strong></font><br />
<BR><br />
<BR><br />
I. Evaluation of TR-HHR<br />
<BR><BR><br />
II. Evaluation of TR-HHR-taRNA<br />
<br />
<BR><BR><BR><BR><br />
<br />
<font size=6 id="TR-HHR"><strong>I. Evaluation of TR-HHR</strong></font><br />
<br />
<BR><br />
<br />
<p style="line-height:110%"><br />
In order to evaluate the regulation of HHR’s self-cleavage activity by taRNA, GFP gene was connected with the HHR or inactive HHR (*HHR), which has a mutation lose the activity.[BBa_K1053004][BBa_K1053005]<BR> These fragments were subcloned into conventional biobrick standard vector pSB3C5. <br />
</p><br />
<br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/4/45/Wiki4.png" width="400px" / ><br />
<BR><br />
<br />
1) <I>E. coli</I> TOP10 strain was co-transformed by two plasmids listed bellow, <br />
<BR><br />
<BR><br />
A)<br />
<BR><br />
pSB1A3-taR12-DT<br />
<BR><br />
pSB3C5-TR(12)-HHR -GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
B)<br />
<BR><br />
pSB1A3-empty<br />
<BR><br />
pSB3C5- TR(12)-HHR -GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
<br />
C) <br />
<BR><br />
pSB1A3-taR12-DT<br />
<BR><br />
pSB3C5-TR(12)- * HHR-GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
<br />
D)<br />
<BR><br />
pSB1A3 empty<br />
<BR><br />
pSB3C5- TR(12)- * HHR -GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
<br />
E) <br />
<BR>pSB1A3-empty<br />
<BR><br />
pSB3C5-empty<br />
<br />
<BR><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
2) Bacterial cultures were incubated in LB medium at 140 rpm and 37℃ for 12 hour. <BR><br />
3) Then, cultures was diluted to an OD595 of 0.0125 and induced with or without 0.1% arabinose.<BR><br />
4) Cultures were incubated with shaking at 140 rpm and 37℃ for 8 hours in 1 mL of LB medium.<BR><br />
5) OD595 and GFP fluorescence intensity were measured.<BR><br />
All experiments were performed using three cultures per each sample.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
Result and Discussion<br />
<br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/f/fb/%E3%82%B9%E3%83%A9%E3%82%A4%E3%83%897.jpg width="400px"><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
In the presence of taR12, GFP fluorescence value divided by OD value is higher than one in absence of taR12. This result was not expected. HHR has the ability to self-cleave, but its ability is suppressed when taRNA hybridizes to HHR. Therefore, in the presence of taRNA, self-cleavage of HHR should not happen. So then, why we get such result? One factor can be due to the exposure of RBS sequence caused by the structural change of HHR when it hybridizes to taRNA<br />
</p><br />
<br />
<BR><BR><BR><BR><br />
<br />
<font size=6 id="TR-HHR-taRNA">II.Evaluation of TR-HHR-taRNA</font><br />
<br />
<BR><BR><br />
<br />
We examined whether TR-HHR-taRNA’s output can be repressed by corresponding taRNA. <br />
<BR><br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/5/52/Wiki5.png" width="400px" / ><br />
<BR><br />
<br />
<br />
1) The plasmids as shown below was used to transform <I>E. coli </I>Top10.<BR><br />
<BR><br />
A) <br />
<BR><br />
pSB1A3-P<sub>bad</sub> -TR(42)HHR-taR12-DT<br />
<BR><br />
pSB3C5-crR12-GFP<br />
<br />
<BR><BR><BR><br />
<br />
B) <br />
<BR><br />
pSB1A3-P<sub>const(H)</sub>-taR42-P<sub>bad</sub>-TR(42)HHR-taR12-DT<BR><br />
pSB3C5-crR12-GFP-DT<br />
<br />
<BR><BR><BR><br />
<br />
C)<br />
<BR><br />
pSB1A3-P<sub>bad</sub>-taR12-DT<BR><br />
pSB3C5-crR12-GFP<br />
<br />
<BR><BR><BR><br />
<br />
D) <BR><br />
pSB1A3-empty <BR><br />
pSB3C5-crR12-GFP<br />
<br />
<BR><BR><BR><br />
<br />
E) <BR><br />
pSB1A3-empty<BR><br />
pSB3C5-empty<BR><br />
<br />
<BR><BR><br />
<br />
<p style="line-height:110%"><br />
2) The transformants were pre-cultured in LB medium at 37 degrees celcius for 8 hours. <BR><br />
3) Then, cultures was diluted to an OD595 of 0.0125 and induced with or without 0.1% arabinose.<BR><br />
4) Cultures were incubated with shaking at 140 rpm and 37℃ for 8 hours in 1 mL of LB medium.<BR><br />
5) OD595 and GFP fluorescence intensity were measured.<BR><br />
</p><br />
<br />
<BR><BR><br />
<br />
<font size=5 ><strong>Result and Discussion</strong></font><br />
<br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/c/c3/%E5%AE%9F%E9%A8%93%EF%BC%92%E7%B5%90%E6%9E%9C%EF%BC%91.jpg width="400px"><br />
<BR><br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/6/69/%E5%AE%9F%E9%A8%93%EF%BC%92%E7%B5%90%E6%9E%9C%EF%BC%92.jpg width="400px"><br />
<BR><br />
<p style="line-height:110%"><br />
In order to evaluate a function of TR(42)-HHR-taR12, we used crR12-GFP as a reporter.<br />
taRNA is complementary to the sequence of crRNA, and by the binding of taRNA to crRNA, RBS is exposed and gene expression is activated. In this result, the GFP fluorescence value divided by OD value of (2) was lower than that of (3). It shows that taR42 suppressed self-cleavage of TR(42)-HHR and caused less GFP expression. Therefore, we prove that the self-cleavage activity of TR(42)-HHR and amount of taR12 can be controlled by taR42.<br />
</p><br />
<br />
<BR><BR><BR><BR><br />
<br />
<font size=6 id="Future work" ><strong>・Future work</strong></font><br />
<br />
<BR><br />
We must confirm whether taR12 and TR(12)-HHR set is available for RNA oscillator.<BR><br />
The second experiment result showed high background, then we should try to use lower promoter to transcribe crR12-GFP.<BR><br />
<BR><br />
<br />
<font size=6 ><strong>Reference</strong></font><br />
<br />
<BR><BR><br />
<BR><br />
<p style="line-height:110%"><br />
[1]<br />
An engineered small RNA-mediated genetic switch<br />
based on a ribozyme expression platform<br />
Benedikt Klauser and Jo¨ rg S. Hartig <br />
Nucleic Acids Research, 2013<br />
<BR><br />
[2]<br />
Callura,J.M., Cantor,C.R. and Collins,J.J. (2012) Genetic<br />
switchboard for synthetic biology applications. Proc. Natl Acad.<br />
Sci. USA, 109, 5850–5855<br />
</p><br />
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<BR><br />
<br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/oscillator
Team:Tokyo-NoKoGen/oscillator
2013-09-28T04:04:54Z
<p>Jinhee: </p>
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<title>Team:Tokyo-NoKoGen - 2013.igem.org</title><br />
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<h1 id="index_title"></h1><br />
<ul id="contents"><br />
<li><a href="#Introduction"><strong>Introduction</strong></a></li><br />
<li><a href="#Objective"><strong>Objective</strong></a></li><br />
<li><a href="#Method"><strong>Method</strong></a><br />
<ul><br />
<li><a href="#Design ">Design </a></li><br />
<li><a href="#Parts construction">Parts construction</a></li><br />
</ul><br />
</li> <br />
<li><a href="#Evaluation"><strong>Evaluation</strong></a><br />
<ul><br />
<li><a href="#TR-HHR">TR-HHR</a></li><br />
<li><a href="#TR-HHR-taRNA">TR-HHR-taRNA</a></li><br />
</ul><br />
</li><br />
<li><a href="#Future work"><strong>Future work</strong></a></li><br />
<br />
</ul><br />
<br />
<ul id="contents2"><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/oscillator"><li>RNA oscillator</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/scaffold"><li>RNAScaffold</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/light"><li>Light sensor</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/modeling"><li>Modeling</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/rhodopsin"><li>Improving a BioBrick part - Rhodopsin</li></a><br />
</ul><br />
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<div id="main"><br />
<br />
<font size=5><br />
<BR><br />
<p align=center><font size=7><strong>RNA Oscillator</strong></font><hr></p><br />
<BR><br />
<BR><br />
<BR><br />
<font size=6><strong>Introduction </span></strong></font><br />
<BR><br />
<BR><br />
<BR><br />
<font size=6 ><strong>Protein oscillator</span></strong></font><br />
<BR><br />
<BR><br />
<p style="line-height:110%"><br />
<BR><br />
The oscillating network was built by using three transcriptional repressor systems in which the first repressor protein suppresses the expression of second repressor gene, whose protein suppresses the transcription of the third gene, and the protein the third gene code suppresses the first gene expression.<br />
</p><br />
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<BR><br />
<BR><br />
<br />
<font size=6><strong>Hummer headed ribozyme</span></strong></font><br />
<br />
<BR><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
Hummerheaded ribozymes (HHRs) are found in many species and known to have a capability to self-cleave. Recently, engineered HHRs responding to trans-acting RNAs (taRNAs) in <I>E.coli</I> were reported. In absence of taRNA, the HHRs can self-cleave, but in presence of taRNA, their capability of self-cleavage is inhibited due to hybridize with taRNA and change their structures.<br />
</p><br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/3/3a/%E3%82%AD%E3%83%A3%E3%83%97%E3%83%81%E3%83%A318.PNG><BR><br />
<br />
<BR><BR><BR><br />
<br />
<p style="line-height:110%"><br />
We developed the new oscillator system that is composed only RNAs not using repressor gene and protein. We created three taRNA connected hummerheaded ribozymes (HHR-taRNA) that sense to another taRNA. The same way as the protein oscillator, three HHR-taRNAs inhibits other one’s self-cleavage such as three-way standoff. When the first HHR-taRNA is self-cleaved, connected the first taRNA that binds to the second HHR-taRNA is free and suppresses the second HHR-taRNA self-cleavage. When second HHR-taRNA is self-cleaved, the second taRNA that binds to the third HHR-taRNA is free and suppresses the third HHR-taRNA. Finally, when third HHR-taRNA is self-cleaved, the third taRNA that binds to the first HHR-taRNA is free and suppresses the first HHR-taRNA, completing cycle.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
<font size=6 id="Objective"><strong>Objective</span></strong></font><br />
<BR><br />
<BR><br />
Construction of Three of orthogonal taRNA generator which repressed by another taRNA.<br />
<br />
<BR><br />
<br />
<p style="line-height:110%"><br />
taRNA responsive HHR (TR-HHR) was fused with taRNA. By using orthogonal riboregulator sequences, we designed three of taRNA generators.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
<font size=6 id="Method"><strong>Method</span></strong></font><br />
<br />
<BR><BR><br />
<br />
<font size=5 id="Design"><strong>-Design</span></strong></font> <br />
<br />
<BR><BR><br />
<br />
<p style="line-height:110%"><br />
TR-HHR was designed by Benedikt Klauser and Jo¨ rg S. Hartig. Their TR-HHR were inspired by the riboregulatory system previously reported by Collins and coworkers. They made use of sequence domains of the riboregulator RR12. And, orthogonal variants of riboregulator RR12 (RR12y, RR42) were reported by Collins and coworkers. So, by using the sequences of these orthogonal riboregulators, we designed three orthogonal TR-HHRs.<br />
</p><br />
<br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/a/a1/Rrvariants.png" width="400px" / ><br />
<img src="https://static.igem.org/mediawiki/2013/c/ce/Cccc.png" width="200px" / ><br />
<br />
<br />
<BR><BR><br />
<br />
<br />
<BR><BR><BR><br />
<br />
<font size=5 id="Parts construction"><strong>-Parts construction</span></strong></font> <br />
<BR><br />
<p style="line-height:110%"><br />
1. Construction of TR-HHR<BR><br />
2. Construction of TR-HHR variants<BR><br />
3. Construction of taRNA variants<BR><br />
4. Construction of TR-HHR-taRNA<BR><br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
1. Construction of TR-HHR<BR><br />
<br />
<p style="line-height:110%"> <br />
We used the TR-HHR developed by Klauser <I>et al.</I>[1] Two overlap primers designed, and PCR was performed to connect them. The PCR product was ligated into pSB1C3 vector.<br />
</p><br />
<br />
<BR><br />
<br />
<img src="https://static.igem.org/mediawiki/igem.org/e/e3/Wiki1.png" width="400px" / ><br />
<br />
<BR><BR><br />
<br />
2.Construction of TR-HHR variants<br />
<br />
<BR><BR><BR><br />
<br />
<p style="line-height:110%"><br />
By inverse PCR using pSB1C3-TR-HHR as a template, we could obtain three TR-HHRs, TR(12)-HHR, TR(12y)-HHR, and TR(42)-HHR.<br />
</p><br />
<br />
<img src="https://static.igem.org/mediawiki/igem.org/9/98/Wiki6.png" width="400px" / ><br />
<br />
<BR><br />
<BR><br />
<BR><br />
<br />
<br />
3.Construction of taRNA variants<br />
<br />
<BR><br />
<BR><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
By inverse PCR using pSB1A3-taR12-DT as a template, we could obtain three taRNAs, taR12, taR12y, taR42.<br />
</p><br />
<br />
<img src="https://static.igem.org/mediawiki/igem.org/a/a7/Wiki2.png" width="400px" / ><br />
<br />
<BR><br />
<BR><br />
<BR><br />
<br />
4.Construction of TR-HHR-taRNA<br />
<BR><br />
<BR><br />
<BR><br />
<br />
TR-HHR-taRNAs were constructed by overlap PCR.<br />
<br />
<BR><br />
<BR><br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/9/91/Wiki3.png" width="400px" / ><br />
<br />
<BR><br />
<BR><br />
<BR><br />
<BR><br />
<BR><br />
<BR><br />
<font size=6 id="Evaluation"><strong>Evaluation</strong></font><br />
<BR><br />
<BR><br />
I. Evaluation of TR-HHR<br />
<BR><BR><br />
II. Evaluation of TR-HHR-taRNA<br />
<br />
<BR><BR><BR><BR><br />
<br />
<font size=6 id="TR-HHR"><strong>I. Evaluation of TR-HHR</strong></font><br />
<br />
<BR><br />
<br />
<p style="line-height:110%"><br />
In order to evaluate the regulation of HHR’s self-cleavage activity by taRNA, GFP gene was connected with the HHR or inactive HHR (*HHR), which has a mutation lose the activity. These fragments were subcloned into conventional biobrick standard vector pSB3C5. <br />
</p><br />
<br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/4/45/Wiki4.png" width="400px" / ><br />
<BR><br />
<br />
1) <I>E. coli</I> TOP10 strain was co-transformed by two plasmids listed bellow, <br />
<BR><br />
<BR><br />
A)<br />
<BR><br />
pSB1A3-taR12-DT<br />
<BR><br />
pSB3C5-TR(12)-HHR -GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
B)<br />
<BR><br />
pSB1A3-empty<br />
<BR><br />
pSB3C5- TR(12)-HHR -GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
<br />
C) <br />
<BR><br />
pSB1A3-taR12-DT<br />
<BR><br />
pSB3C5-TR(12)- * HHR-GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
<br />
D)<br />
<BR><br />
pSB1A3 empty<br />
<BR><br />
pSB3C5- TR(12)- * HHR -GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
<br />
E) <br />
<BR>pSB1A3-empty<br />
<BR><br />
pSB3C5-empty<br />
<br />
<BR><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
2) Bacterial cultures were incubated in LB medium at 140 rpm and 37℃ for 12 hour. <BR><br />
3) Then, cultures was diluted to an OD595 of 0.0125 and induced with or without 0.1% arabinose.<BR><br />
4) Cultures were incubated with shaking at 140 rpm and 37℃ for 8 hours in 1 mL of LB medium.<BR><br />
5) OD595 and GFP fluorescence intensity were measured.<BR><br />
All experiments were performed using three cultures per each sample.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
Result and Discussion<br />
<br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/f/fb/%E3%82%B9%E3%83%A9%E3%82%A4%E3%83%897.jpg width="400px"><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
In the presence of taR12, GFP fluorescence value divided by OD value is higher than one in absence of taR12. This result was not expected. HHR has the ability to self-cleave, but its ability is suppressed when taRNA hybridizes to HHR. Therefore, in the presence of taRNA, self-cleavage of HHR should not happen. So then, why we get such result? One factor can be due to the exposure of RBS sequence caused by the structural change of HHR when it hybridizes to taRNA<br />
</p><br />
<br />
<BR><BR><BR><BR><br />
<br />
<font size=6 id="TR-HHR-taRNA">II.Evaluation of TR-HHR-taRNA</font><br />
<br />
<BR><BR><br />
<br />
We examined whether TR-HHR-taRNA’s output can be repressed by corresponding taRNA. <br />
<BR><br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/5/52/Wiki5.png" width="400px" / ><br />
<BR><br />
<br />
<br />
1) The plasmids as shown below was used to transform <I>E. coli </I>Top10.<BR><br />
<BR><br />
A) <br />
<BR><br />
pSB1A3-P<sub>bad</sub> -TR(42)HHR-taR12-DT<br />
<BR><br />
pSB3C5-crR12-GFP<br />
<br />
<BR><BR><BR><br />
<br />
B) <br />
<BR><br />
pSB1A3-P<sub>const(H)</sub>-taR42-P<sub>bad</sub>-TR(42)HHR-taR12-DT<BR><br />
pSB3C5-crR12-GFP-DT<br />
<br />
<BR><BR><BR><br />
<br />
C)<br />
<BR><br />
pSB1A3-P<sub>bad</sub>-taR12-DT<BR><br />
pSB3C5-crR12-GFP<br />
<br />
<BR><BR><BR><br />
<br />
D) <BR><br />
pSB1A3-empty <BR><br />
pSB3C5-crR12-GFP<br />
<br />
<BR><BR><BR><br />
<br />
E) <BR><br />
pSB1A3-empty<BR><br />
pSB3C5-empty<BR><br />
<br />
<BR><BR><br />
<br />
<p style="line-height:110%"><br />
2) The transformants were pre-cultured in LB medium at 37 degrees celcius for 8 hours. <BR><br />
3) Then, cultures was diluted to an OD595 of 0.0125 and induced with or without 0.1% arabinose.<BR><br />
4) Cultures were incubated with shaking at 140 rpm and 37℃ for 8 hours in 1 mL of LB medium.<BR><br />
5) OD595 and GFP fluorescence intensity were measured.<BR><br />
</p><br />
<br />
<BR><BR><br />
<br />
<font size=5 ><strong>Result and Discussion</strong></font><br />
<br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/c/c3/%E5%AE%9F%E9%A8%93%EF%BC%92%E7%B5%90%E6%9E%9C%EF%BC%91.jpg width="400px"><br />
<BR><br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/6/69/%E5%AE%9F%E9%A8%93%EF%BC%92%E7%B5%90%E6%9E%9C%EF%BC%92.jpg width="400px"><br />
<BR><br />
<p style="line-height:110%"><br />
In order to evaluate a function of TR(42)-HHR-taR12, we used crR12-GFP as a reporter.<br />
taRNA is complementary to the sequence of crRNA, and by the binding of taRNA to crRNA, RBS is exposed and gene expression is activated. In this result, the GFP fluorescence value divided by OD value of (2) was lower than that of (3). It shows that taR42 suppressed self-cleavage of TR(42)-HHR and caused less GFP expression. Therefore, we prove that the self-cleavage activity of TR(42)-HHR and amount of taR12 can be controlled by taR42.<br />
</p><br />
<br />
<BR><BR><BR><BR><br />
<br />
<font size=6 id="Future work" ><strong>・Future work</strong></font><br />
<br />
<BR><BR><BR><br />
<br />
<font size=6 ><strong>Reference</strong></font><br />
<br />
<BR><BR><br />
<BR><br />
<p style="line-height:110%"><br />
[1]<br />
An engineered small RNA-mediated genetic switch<br />
based on a ribozyme expression platform<br />
Benedikt Klauser and Jo¨ rg S. Hartig <br />
Nucleic Acids Research, 2013<br />
<BR><br />
[2]<br />
Callura,J.M., Cantor,C.R. and Collins,J.J. (2012) Genetic<br />
switchboard for synthetic biology applications. Proc. Natl Acad.<br />
Sci. USA, 109, 5850–5855<br />
</p><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Daily_notes
Team:Sumbawagen/Daily notes
2013-09-28T04:02:26Z
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<br><br />
Notes for July 17th 2013<br />
<br><br />
Activities:<br><br />
1. First briefing with all members of IGEM (advisors, instructors, student) plus some members’s of University’s officials.<br />
<br><br />
2. Self-intoduction from students members about his/her biodata and motivation.<br />
<br><br />
3. Explanation of IGEM Team Sumbawagen schedule, and Human Practice project from Dr. Arief.<br />
<br><br />
4. Election of student leader by students themselves.<br />
<br><br />
<br />
Picture :<br><br />
We are sorry, we have no pictures for the first meeting, because we were too focus for the first meeting.<br />
<br><br />
<a href="http://photozou.jp/photo/show/2832099/188348829"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188348829.png" alt="スクリーンショット 2013-09-28 12.58.18" width="450" height="438"></a><br><a href="http://photozou.jp/photo/show/2832099/188348829"></a> <br><br />
<a href="http://photozou.jp/photo/show/2832099/188348841"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188348841.png" alt="スクリーンショット 2013-09-28 12.58.28" width="450" height="337"></a><br><a href="http://photozou.jp/photo/show/2832099/188348841"></a> <br><br />
<a href="http://photozou.jp/photo/show/2832099/188348848"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188348848.png" alt="スクリーンショット 2013-09-28 12.58.39" width="450" height="181"></a><br><a href="http://photozou.jp/photo/show/2832099/188348848"></a> <br><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Daily_notes
Team:Sumbawagen/Daily notes
2013-09-28T04:01:55Z
<p>Jinhee: </p>
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<br><br />
Notes for July 17th 2013<br />
<br><br />
Activities:<br><br />
1. First briefing with all members of IGEM (advisors, instructors, student) plus some members’s of University’s officials.<br />
<br><br />
2. Self-intoduction from students members about his/her biodata and motivation.<br />
<br><br />
3. Explanation of IGEM Team Sumbawagen schedule, and Human Practice project from Dr. Arief.<br />
<br><br />
4. Election of student leader by students themselves.<br />
<br><br />
<br />
Picture :<br><br />
We are sorry, we have no pictures for the first meeting, because we were too focus for the first meeting.<br />
<br><br />
<a href="http://photozou.jp/photo/show/2832099/188348829"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188348829.png" alt="スクリーンショット 2013-09-28 12.58.18" width="450" height="438"></a><br><a href="http://photozou.jp/photo/show/2832099/188348829">スクリーンショット 2013-09-28 12.58.18</a> <br><br />
<a href="http://photozou.jp/photo/show/2832099/188348841"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188348841.png" alt="スクリーンショット 2013-09-28 12.58.28" width="450" height="337"></a><br><a href="http://photozou.jp/photo/show/2832099/188348841">スクリーンショット 2013-09-28 12.58.28</a> <br><br />
<a href="http://photozou.jp/photo/show/2832099/188348848"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188348848.png" alt="スクリーンショット 2013-09-28 12.58.39" width="450" height="181"></a><br><a href="http://photozou.jp/photo/show/2832099/188348848">スクリーンショット 2013-09-28 12.58.39</a> <br><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Daily_notes
Team:Sumbawagen/Daily notes
2013-09-28T04:01:06Z
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<br><br />
Notes for July 17th 2013<br />
<br><br />
Activities:<br><br />
1. First briefing with all members of IGEM (advisors, instructors, student) plus some members’s of University’s officials.<br />
<br><br />
2. Self-intoduction from students members about his/her biodata and motivation.<br />
<br><br />
3. Explanation of IGEM Team Sumbawagen schedule, and Human Practice project from Dr. Arief.<br />
<br><br />
4. Election of student leader by students themselves.<br />
<br><br />
<br />
Picture :<br><br />
We are sorry, we have no pictures for the first meeting, because we were too focus for the first meeting.<br />
<br><br />
<a href="http://photozou.jp/photo/show/2832099/188348829"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188348829.png" alt="スクリーンショット 2013-09-28 12.58.18" width="450" height="438"></a><br><a href="http://photozou.jp/photo/show/2832099/188348829">スクリーンショット 2013-09-28 12.58.18</a> <br><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Daily_notes
Team:Sumbawagen/Daily notes
2013-09-28T03:57:39Z
<p>Jinhee: </p>
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<br><br />
Notes for July 17th 2013<br />
<br><br />
Activities:<br><br />
1. First briefing with all members of IGEM (advisors, instructors, student) plus some members’s of University’s officials.<br />
<br><br />
2. Self-intoduction from students members about his/her biodata and motivation.<br />
<br><br />
3. Explanation of IGEM Team Sumbawagen schedule, and Human Practice project from Dr. Arief.<br />
<br><br />
4. Election of student leader by students themselves.<br />
<br><br />
<br />
Picture :<br><br />
We are sorry, we have no pictures for the first meeting, because we were too focus for the first meeting.<br />
<br><br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/questionnaire
Team:Tokyo-NoKoGen/questionnaire
2013-09-28T03:50:17Z
<p>Jinhee: </p>
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<a href="https://2013.igem.org"><img class="iGEM" src="https://static.igem.org/mediawiki/2013/5/58/IGEM.png"></a> <br />
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<div id="mymenubar"><br />
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<table><br />
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<td class="home"><a href="https://2013.igem.org/Team:Tokyo-NoKoGen"><span class="home"></span></a></td><br />
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<td class="project"><a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Project"><span class="project"></span></a></td><br />
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<div id="index"><br />
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<h1 id="index_title"></h1><br />
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<br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/labosafety"><li><strong>Safety in the laboratoy</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/questionnaire"><li><strong>Questionnaire on synthetic biology</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/ikalux"><li><strong>Do It Yourself -Ika Lux-</strong></li></a><br />
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<div id="main"><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=6>Questionnaire on synthetic biology</font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>We would like to thank all the people who helped us answer the questionnaire, we were able to get 85 replies by 2 p.m. on 27th of September. We prepared 21 questions to find out how people recognize synthetic biology. </p></font><br />
<br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q1. Are you a male or a female?</font></p><br />
<p align=center><font size=3>性別を教えてください</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/5/52/%E3%82%B9%E3%83%A9%E3%82%A4%E3%83%891.JPG></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>The ratio is almost ideal!</font></p><br />
<br />
<BR><br />
<p align=center><font size=4>Q2. What age are you?</font></p><br />
<p align=center><font size=3>年代を教えてください</p></font><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/1/1f/Q2.JPG></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Unfortunately, the ratio of generations is biased. This means that this questionnaire tend to reflect younger people's opinion.</font></p><br />
<BR><br />
<p align=center><font size=4>Q3. Are you specialized in science or non-science?</font></p><br />
<p align=center><font size=3>文系か理系、どちらがお得意かお答えください</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/2/2c/Q3.JPG></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Most of people are major in science. </font></p><br />
<BR><br />
<p align=center><font size=4>Q4. Do you know synthetic biology?</font></p><br />
<p align=center><font size=3>「合成生物学」を知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/5/52/Q4.JPG></p><br />
<BR><br />
<p align=center><font size=4> </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Even though many people who involved in this questionnaire are major in science, it seems that the word "Synthetic biology" is unfamiliar.</font></p><br />
<BR><br />
<p align=center><font size=4>Q5. If yes, how did you get to know about it?</font></p><br />
<p align=center><font size=3> 「はい」と答えた方はどこで知りましたか?<br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/0/03/5.jpg width=600></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Most of people who know Synthetic biology get the information about synthetic biology from their friends .</font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q6. What do you think Synthetic biology is about?</font></p><br />
<p align=center><font size=3> 合成生物学は次のうちどのような学問だと思いますか?<br />
<p alignt=center><img src=https://static.igem.org/mediawiki/2013/f/f9/Q6.JPG width=600></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>When people are asked “what do you think Synthetic biology is about?”, most of people have right recognition about synthetic biology. </font></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q7. Do you know iGEM?</font></p><br />
<p align=center><font size=3>iGEMという大会を知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/1/11/Q7.JPG></p><br />
<BR><br />
<p align=center><font size=4>About 60% of people don’t know iGEM. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q8. If yes, how do you know it?</font></p><br />
<p align=center><font size=3>「はい」と答えた方はどこで知りましたか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/9/9f/Q8.JPG></p><br />
<BR><br />
<p align=center><font size=4>Most of people who know iGEM, they knew the iGEM from their friend as well as Q5. So, people who know synthetic biology probably know iGEM. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q9. How do you pronounce "iGEM"?</font></p><br />
<p align=center><font size=3>"iGEM"の読み方は次のうちどれだと思いますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/0/04/Q9.JPG></p><br />
<BR><br />
<p align=center><font size=4>About 60% people know right pronunciation of iGEM. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>10. Where do you think the iGEM Jamboree is held?</font></p><br />
<p align=center><font size=3>iGEMの本選はどこで開催されていると思いますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/1/19/Q10.JPG></p><br />
<BR><br />
<p align=center><font size=4>About 50% of people don’t know where iGEM Jamboree held. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q11. What do you think is important in iGEM?</font></p><br />
<p align=center><font size=3>iGEMとはどのようなことを競う大会だと思いますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/4/4b/Q11.JPG width=700></p><br />
<BR><br />
<p align=center><font size=4>回答は三パターンにわかれた. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q12. How many Japanese teams do you think participate iGEM?</font></p><br />
<p align=center><font size=3>iGEMの日本のチームは何チームいるか知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/e/e1/Q12.JPG></p><br />
<BR><br />
<p align=center><font size=4>About 50% of people recognize the right number of Japanese teams take part in iGEM. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q13. How many teams do you think there are in the whole iGEM society?</font></p><br />
<p align=center><font size=3>世界にはiGEMが何チームいるか知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/8/8f/Q13.JPG></P><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q14. What do you think of, when you hear "E. coli"?</font></p><br />
<p align=center><font size=3>「大腸菌」という単語から連想するものは何ですか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/c/cf/Q14.JPG></P><br />
<BR><br />
<p align=center><font size=4>Most answered question is the Good for science. But the second is Dirty. People who specialize in science can recognize E.coli was the Good for science. But common people tend to imagine that E.coli is dirty. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q15. What do you think of, when you hear "Genetic modification"?</font></p><br />
<p align=center><font size=3>「遺伝子組み換え」という単語から連想するものは何ですか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/4/4b/Q15.JPG></p><br />
<BR><br />
<p align=center><font size=4>Most answered question is advenced technology but the second is "scary". </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q16. Did you know that genetically modified E. coli can collect heavy metal ions?</font></p><br />
<p align=center><font size=3>遺伝子組み換え型大腸菌を使って重金属を回収することが出来ることを知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/a/a0/Q16.JPG></p><br />
<BR><br />
<p align=center><font size=4>About 70% of people don’t know that modified E.coli can collect heavy metal ions. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q17. Did you know that genetically modified E. coli can become luminescent?</font></p><br />
<p align=center><font size=3>遺伝子組み換え型大腸菌が青色に光ることが出来ることを知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/4/40/Q17.JPG></p><br />
<BR><br />
<p align=center><font size=4>About 70% of people don’t know that modified E.coli can become luminescent. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q18. Did you know that genetically modified E. coli can be regulated by light?</font></p><br />
<p align=center><font size=3>遺伝子組み換え型大腸菌を光で制御出来ることを知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/2/22/Q18.JPG></p><br />
<BR><br />
<p align=center><font size=4>About 70% of people don’t know that modified E.coli can can be regulated by light. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q19. We bought a squid at a nearby supermarket. What do you think we did with them?</font></p><br />
<p align=center><font size=3>私達は近所のスーパーで石川県産のイカを買いました。そのイカをどうしたでしょう。</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/e/e6/Q19.JPG></p><br />
<BR><br />
<p align=center><font size=4>About 50 % of people imagine correctly what we have done with a squid. And this is the most important question because people who know the right answer recognize that we can enjoy synthetic biology using familiar things around everyday life. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q20. After this questionnaire, do you think synthetic biology is interesting?</font></p><br />
<p align=center><font size=3>合成生物学について、なんとなく興味を持っていただけたでしょうか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/d/db/Q20.JPG></p><br />
<BR><br />
<p align=center><font size=4>Most of people became interested in synthetic biology via this questionarier. So, we were succeeded to attract people to synthetic biology. </font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q21. Do you also feel that you want to participate iGEM competitiong?</font></p><br />
<p align=center><font size=3>iGEMに参加したいと思ってもらえたでしょうか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/5/5a/Q21.JPG></p><br />
<BR><br />
<p align=center><font size=4>As the results of Q20 and Q21, most of people became interested in synthetic biology but about 60% of people don’t feel like to participate iGEM competiting. This is because awareness of iGEM is still low. So when people recognize iGEM is very interesting competition, more people can feel like to participate in iGEM!!. </font></p><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Notebook
Team:Sumbawagen/Notebook
2013-09-28T03:46:18Z
<p>Jinhee: </p>
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<a href="http://photozou.jp/photo/show/2832099/188347227"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347227.png" alt="Fahmi explained about transformation process" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347227">Fahmi explained about transformation process</a> <br />
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<a href="http://photozou.jp/photo/show/2832099/188347234"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347234.png" alt="Honey glucose measurement" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347234">Honey glucose measurement</a><br />
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<a href="http://photozou.jp/photo/show/2832099/188347245"><img src="http://kura3.photozou.jp/pub/99/2832099/photo/188347245.png" alt="Pouring medium into petri dishes" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347245">Pouring medium into petri dishes</a><br />
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<a href="http://photozou.jp/photo/show/2832099/188347255"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347255.jpg" alt="Prof. Sode comments after Sumbawagen iGEM Team presentation" width="450" height="338"></a><br><a href="http://photozou.jp/photo/show/2832099/188347255">Prof. Sode comments after Sumbawagen iGEM Team presentation</a> <br />
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<a href="http://photozou.jp/photo/show/2832099/188347264"><img src="http://kura3.photozou.jp/pub/99/2832099/photo/188347264.png" alt="Study" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347264">Study</a> <br />
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<a href="http://photozou.jp/photo/show/2832099/188347275"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347275.jpg" alt="Sumbawagen team after presentation" width="450" height="338"></a><br><a href="http://photozou.jp/photo/show/2832099/188347275">Sumbawagen team after presentation</a><br />
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<a href="http://photozou.jp/photo/show/2832099/188347289"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347289.png" alt="Sumbawagen team and Prof. Koji Sode after seminar" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347289">Sumbawagen team and Prof. Koji Sode after seminar</a> <br />
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<a href="http://photozou.jp/photo/show/2832099/188347301"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347301.png" alt="Sumbawagen team in lab" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347301">Sumbawagen team in lab</a> <br />
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<a href="http://photozou.jp/photo/show/2832099/188347318"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347318.png" alt="Sumbawagen team surveyed in senior high school" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347318">Sumbawagen team surveyed in senior high school</a><br />
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<a href="http://photozou.jp/photo/show/2832099/188347336"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347336.png" alt="Sumbawagen team with Ms. Dwi" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347336">Sumbawagen team with Ms. Dwi</a> <br />
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<a href="http://photozou.jp/photo/show/2832099/188347346"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347346.png" alt="Sumbawagen team with Prof. Koji Sode and Dr. Tomohiko Yamazaki" width="448" height="302"></a><br><a href="http://photozou.jp/photo/show/2832099/188347346">Sumbawagen team with Prof. Koji Sode and Dr. Tomohiko Yamazaki</a> <br><br />
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<a href="http://photozou.jp/photo/show/2832099/188347370"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347370.png" alt="Sumbawagen team with SUT staffs" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347370">Sumbawagen team with SUT staffs</a><br><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Notebook
Team:Sumbawagen/Notebook
2013-09-28T03:45:15Z
<p>Jinhee: </p>
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<br><br />
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<a href="http://photozou.jp/photo/show/2832099/188347275"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347275.jpg" alt="Sumbawagen team after presentation" width="450" height="338"></a><br><a href="http://photozou.jp/photo/show/2832099/188347275">Sumbawagen team after presentation</a><br />
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<a href="http://photozou.jp/photo/show/2832099/188347318"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347318.png" alt="Sumbawagen team surveyed in senior high school" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347318">Sumbawagen team surveyed in senior high school</a><br />
<br><br />
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<a href="http://photozou.jp/photo/show/2832099/188347336"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347336.png" alt="Sumbawagen team with Ms. Dwi" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347336">Sumbawagen team with Ms. Dwi</a> <br />
<br><br />
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<a href="http://photozou.jp/photo/show/2832099/188347346"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347346.png" alt="Sumbawagen team with Prof. Koji Sode and Dr. Tomohiko Yamazaki" width="448" height="302"></a><br><a href="http://photozou.jp/photo/show/2832099/188347346">Sumbawagen team with Prof. Koji Sode and Dr. Tomohiko Yamazaki</a> <br><br />
<br />
<a href="http://photozou.jp/photo/show/2832099/188347356"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347356.png" alt="Sumbawagen team with SUT staffs-3" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347356">Sumbawagen team with SUT staffs-3</a><br><br />
<br />
<a href="http://photozou.jp/photo/show/2832099/188347370"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347370.png" alt="Sumbawagen team with SUT staffs" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347370">Sumbawagen team with SUT staffs</a><br><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Notebook
Team:Sumbawagen/Notebook
2013-09-28T03:42:59Z
<p>Jinhee: </p>
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<br />
<a href="http://photozou.jp/photo/show/2832099/188347255"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347255.jpg" alt="Prof. Sode comments after Sumbawagen iGEM Team presentation" width="450" height="338"></a><br><a href="http://photozou.jp/photo/show/2832099/188347255">Prof. Sode comments after Sumbawagen iGEM Team presentation</a> <br />
<br><br />
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<a href="http://photozou.jp/photo/show/2832099/188347264"><img src="http://kura3.photozou.jp/pub/99/2832099/photo/188347264.png" alt="Study" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347264">Study</a> <br />
<br><br />
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<a href="http://photozou.jp/photo/show/2832099/188347275"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347275.jpg" alt="Sumbawagen team after presentation" width="450" height="338"></a><br><a href="http://photozou.jp/photo/show/2832099/188347275">Sumbawagen team after presentation</a><br />
<br><br />
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<a href="http://photozou.jp/photo/show/2832099/188347289"><img src="http://kura1.photozou.jp/pub/99/2832099/photo/188347289.png" alt="Sumbawagen team and Prof. Koji Sode after seminar" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347289">Sumbawagen team and Prof. Koji Sode after seminar</a> <br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Notebook
Team:Sumbawagen/Notebook
2013-09-28T03:39:46Z
<p>Jinhee: </p>
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<a href="http://photozou.jp/photo/show/2832099/188347227"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347227.png" alt="Fahmi explained about transformation process-3" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347227">Fahmi explained about transformation process-3</a> <br />
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<a href="http://photozou.jp/photo/show/2832099/188347234"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347234.png" alt="Honey glucose measurement" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347234">Honey glucose measurement</a><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Notebook
Team:Sumbawagen/Notebook
2013-09-28T03:39:16Z
<p>Jinhee: </p>
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#footer-box {<br />
width: 1410px;<br />
height: ;<br />
}<br />
<br />
#footer {<br />
border: none;<br />
width: 1410px;<br />
}<br />
<br />
.firstHeading {<br />
display: none;<br />
}<br />
<br />
#f-list a {<br />
color: #333;<br />
font-size: 10px;<br />
}<br />
<br />
#f-list a:hover {<br />
color: #666;<br />
}<br />
<br />
.printfooter {<br />
display: none;<br />
}<br />
<br />
h3#siteSub {<br />
display: none;<br />
}<br />
<br />
#contentSub {<br />
display: none;<br />
}<br />
<br />
<br />
#menu{ <br />
<br />
width:400px;<br />
<br />
}<br />
<br />
<br />
<br />
</style><br />
<br />
<br />
</head><br />
<body style="background: rgb(255, 255, 255) url(https://static.igem.org/mediawiki/2013/4/4b/Playtime_background_yellow.jpg) repeat scroll top left; margin: 0pt; " onload="onPageLoad();" onunload="onPageUnload();"><br />
<div style="text-align: center; "><br />
<div style="margin-bottom: 15px; margin-left: auto; margin-right: auto; margin-top: 15px; overflow: hidden; position: relative; word-wrap: break-word; text-align: left; width: 700px; " id="body_content"><br />
<div style="background: transparent url(https://static.igem.org/mediawiki/2013/6/65/Playtime_page_bg-6.png) repeat scroll top left; width: 700px; "><br />
<div style="height: 111px; margin-left: 0px; position: relative; width: 700px; z-index: 10; " id="header_layer"><br />
<div style="height: 0px; line-height: 0px; " class="bumper"> </div><br />
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<div class="text-content style_External_645_61" style="padding: 0px; "><br />
<div class="style"><br />
<p style="padding-bottom: 0pt; padding-top: 0pt; " class="paragraph_style">Sumbawagen iGEM Team</p><br />
</div><br />
</div><br />
</div><br />
<br />
<br />
<br />
<div style="height: 22px; width: 645px; height: 22px; left: 22px; position: absolute; top: 29px; width: 645px; z-index: 1; " class="tinyText style_SkipStroke_1"><br />
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<img src="https://static.igem.org/mediawiki/2013/4/40/Kids_green_header_02_1.png" alt="" style="border: none; height: 86px; width: 645px; " /><br />
</div><br />
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<div style="height: 20px; width: 700px; height: 20px; left: 0px; position: absolute; top: 0px; width: 700px; z-index: 1; " class="tinyText style_SkipStroke_1"><br />
<img src="https://static.igem.org/mediawiki/2013/3/36/Playtime_header_bg.jpg" alt="" style="border: none; height: 20px; width: 700px; " /><br />
</div><br />
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<div style="height: 172px; width: 190px; height: 172px; left: 495px; position: absolute; top: 15px; width: 190px; z-index: 1; " class="tinyText style_SkipStroke_1"><br />
<img src="https://static.igem.org/mediawiki/2013/3/3a/Playtime_Sun-4.png" alt="" style="border: none; height: 172px; width: 190px; " /><br />
</div><br />
<div style="height: 80px; width: 80px; height: 80px; left: 10px; position: absolute; top: 15px; width: 190px; z-index: 1; " class="tinyText style_SkipStroke_1"><br />
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</a></div><br />
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</div><br />
<div style="margin-left: 0px; position: relative; width: 700px; z-index: 0; " id="nav_layer"><br />
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<div style="height: 1px; line-height: 1px; " class="tinyText"> </div><br />
<div class="com-apple-iweb-widget-navbar flowDefining" id="widget0" style="margin-left: 22px; margin-top: -1px; opacity: 1.00; position: relative; width: 645px; z-index: 1; "><br />
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<div id="widget0-navbar" class="navbar"><br />
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<div id="widget0-bg" class="navbar-bg"><br />
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<ul id="widget0-navbar-list" class="navbar-list"><br />
<li></li> <br />
</ul><br />
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</div><br />
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<div id="menu"><br />
<A Href="https://2013.igem.org/Team:Sumbawagen">Home</A><br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Team">Team</A><br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Modeling">Modeling</A><br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Project">Project</A><br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Project">Parts</A><br />
Notebook<br />
<br><br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Lab Preparation team activities">Lab Preparation team activities</A><br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Safety">Safety</A><br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Acknowledgement">Acknowledgement</A><br />
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<div style="clear: both; height: 0px; line-height: 0px; " class="spacer"> </div><br />
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<div style="margin-left: 0px; position: relative; width: 700px; z-index: 5; " id="body_layer"><br />
<div style="height: 0px; line-height: 0px; " class="bumper"> </div><br />
<div id="id2" style="height: 34px; left: 35px; position: absolute; top: 17px; width: 620px; z-index: 1; " class="style_SkipStroke_2 shape-with-text"><br />
<div class="text-content Normal_External_620_34" style="padding: 0px; "><br />
<div class="Normal"><br />
<br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Protocols">Protocols</A><br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Artwork_pictures">Artwork pictures</A><br />
<A Href="https://2013.igem.org/Team:Sumbawagen/Daily_notes">Daily notes</A><br />
</div><br />
</div><br />
</div><br />
<br />
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<div style="height: 16px; width: 645px; height: 16px; left: 22px; position: absolute; top: 0px; width: 645px; z-index: 1; " class="tinyText style_SkipStroke_1"><br />
<img src="https://static.igem.org/mediawiki/2013/3/3a/Kids_green_header_03.png" alt="" style="border: none; height: 16px; width: 645px; " /><br />
</div><br />
<br />
<br />
<br />
<div id="id3" style="height: 2766px; left: 35px; position: absolute; top: 47px; width: 628px; z-index: 1; " class="style_SkipStroke_3 shape-with-text"><br />
<div class="text-content graphic_textbox_layout_style_default_External_628_2766" style="padding: 0px; "><br />
<div class="graphic_textbox_layout_style_default"><br />
<br />
<br><br />
<br><br />
<br />
<a href="http://photozou.jp/photo/show/2832099/188347227"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347227.png" alt="Fahmi explained about transformation process-3" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347227">Fahmi explained about transformation process-3</a> <br />
<br />
<br><br />
<br />
<a href="http://photozou.jp/photo/show/2832099/188347234"><img src="http://kura2.photozou.jp/pub/99/2832099/photo/188347234.png" alt="Honey glucose measurement" width="448" height="336"></a><br><a href="http://photozou.jp/photo/show/2832099/188347234">Honey glucose measurement</a><br />
</div><br />
<div style="clear: both; height: 0px; line-height: 0px; " class="tinyText"> </div><br />
</div><br />
</div><br />
<div style="height: 400px; line-height: 400px; " class="spacer"> </div><br />
</div><br />
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<div style="height: 0px; line-height: 0px; " class="bumper"> </div><br />
<br />
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</a><br />
<div style="height: 17px; width: 700px; height: 17px; left: 0px; position: absolute; top: 58px; width: 700px; z-index: 1; " class="tinyText style_SkipStroke_1"><br />
<img src="https://static.igem.org/mediawiki/2013/b/be/Playtime_footer_bg.jpg" alt="" style="border: none; height: 17px; width: 700px; " /><br />
</div><br />
</div><br />
</div><br />
</div><br />
</div><br />
</body><br />
</html></div>
Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/oscillator
Team:Tokyo-NoKoGen/oscillator
2013-09-28T03:28:24Z
<p>Jinhee: </p>
<hr />
<div><html><br />
<br />
<head><br />
<br />
<style type ='text/css'><br />
<br />
#globalWrapper { <br />
width: 1410px; <br />
height:100%;<br />
}<br />
<br />
#content { <br />
background-color: transparent; <br />
border: none; <br />
padding: 0;<br />
margin: 0; <br />
margin-top: -23px; <br />
width: 100%; <br />
overflow: hidden; <br />
height:100%;<br />
}<br />
<br />
#bodyContent { <br />
border: none; <br />
padding:0; <br />
margin:0; <br />
width:1410px; <br />
height:100%;<br />
}<br />
<br />
#top-section {<br />
height: 18px; <br />
margin: 0px; <br />
margin-top: 5px; <br />
margin-left: auto; <br />
margin-right: auto; <br />
margin-bottom: 0 !important;<br />
padding:0; <br />
border: none; <br />
font-size: 10px;<br />
}<br />
<br />
#p-logo { <br />
overflow:hidden; <br />
display: none;<br />
}<br />
<br />
#search-controls { <br />
display:none; <br />
}<br />
<br />
#footer-box {<br />
width: 1410px; <br />
height: ;<br />
}<br />
<br />
#footer { <br />
border: none; <br />
width: 1410px; <br />
}<br />
<br />
.firstHeading { <br />
display: none;<br />
}<br />
<br />
#f-list a { <br />
color: #333; <br />
font-size: 10px;<br />
}<br />
<br />
#f-list a:hover { <br />
color: #666;<br />
}<br />
<br />
.printfooter { <br />
display: none; <br />
}<br />
<br />
h3#siteSub { <br />
display: none;<br />
}<br />
<br />
#contentSub {<br />
display: none;<br />
}<br />
<br />
p:first-child { <br />
display: none;<br />
}<br />
<br />
h1,h2,h3 {<br />
border: none;<br />
}<br />
<br />
#wrapper {<br />
font-size: 12pt;<br />
font-family: "Comic Sans MS","MS明朝";<br />
width:1410px;<br />
height:100%;<br />
margin:auto ;<br />
}<br />
<br />
#title_logo {<br />
float: left;<br />
width:705px;<br />
height:250px;<br />
background :url("https://static.igem.org/mediawiki/2013/d/d6/Header_index_kiso5.png") ;<br />
}<br />
<br />
#title_logo img.iGEM {<br />
position: relative;<br />
top: 50px;<br />
left: 40px;<br />
width: 150px;<br />
height: auto;<br />
}<br />
<br />
#title_logo img.title {<br />
position: relative;<br />
top:20px;<br />
left:40px;<br />
width:600px;<br />
height:200px;<br />
}<br />
<br />
#mymenubar {<br />
float: left;<br />
width: 705px;<br />
height: 250px;<br />
background: url("https://static.igem.org/mediawiki/2013/d/d6/Header_index_kiso5.png") -705px 0px no-repeat;<br />
}<br />
<br />
#mymenubar table,tbody,tr {<br />
background: transparent;<br />
}<br />
<br />
#mymenubar table td div.home,home1 {<br />
width: 70px;<br />
height: 95px;<br />
}<br />
<br />
#mymenubar table td span.home {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 90px 20px 0px 20px;<br />
background: url("https://static.igem.org/mediawiki/2013/9/96/Home28.png") no-repeat 0px 0px;<br />
}<br />
<br />
#mymenubar table td span.home:hover {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 90px 20px 0px 20px;<br />
background-position: 0px -99px;<br />
}<br />
<br />
<br />
#mymenubar table div.home1 {<br />
width: 70px;<br />
height: 95px;<br />
margin: 92px 20px 0px 20px ;<br />
background: url("https://static.igem.org/mediawiki/2013/1/18/Home26.png") no-repeat 0px 0px ;<br />
background-size: 70px 95px;<br />
}<br />
<br />
#mymenubar table td span.project {<br />
display: block;<br />
width: 70px;<br />
height: 95px;<br />
}<br />
<br />
#mymenubar table td.project {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 88px 20px 0px 0px;<br />
background: url("https://static.igem.org/mediawiki/2013/f/f5/Project28.png") no-repeat 0px 0px;<br />
}<br />
<br />
#mymenubar table td.project:hover {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 88px 20px 0px 0px;<br />
background-position: 0px -99px;<br />
}<br />
<br />
#mymenubar table div.project1 {<br />
width: 70px;<br />
height: 95px;<br />
margin: 92px 20px 0px 0px ;<br />
background: url("https://static.igem.org/mediawiki/2013/f/f9/Project26.png") no-repeat 0px 0px ;<br />
background-size: 70px 95px;<br />
}<br />
<br />
<br />
#mymenubar table td span.team {<br />
display: block;<br />
width: 70px;<br />
height: 95px;<br />
}<br />
<br />
#mymenubar table td.team {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 91px 20px 0px 0px;<br />
background: url("https://static.igem.org/mediawiki/2013/b/ba/Team28.png") no-repeat 0px 0px;<br />
}<br />
<br />
#mymenubar table td.team:hover {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 91px 20px 0px 0px;<br />
background-position: 0px -98px;<br />
}<br />
<br />
#mymenubar table div.team1 {<br />
width: 70px;<br />
height: 95px;<br />
margin: 92px 20px 0px 0px ;<br />
background: url("https://static.igem.org/mediawiki/2013/8/8e/Team26.png") no-repeat 0px 0px ;<br />
background-size: 70px 95px;<br />
}<br />
<br />
<br />
#mymenubar table td span.biobrick {<br />
display: block;<br />
width: 70px;<br />
height: 95px;<br />
}<br />
<br />
#mymenubar table td.biobrick {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 93px 20px 0px -2px;<br />
background: url("https://static.igem.org/mediawiki/2013/9/96/Biobrick28.png") no-repeat 0px 0px;<br />
}<br />
<br />
#mymenubar table td.biobrick:hover {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 93px 20px 0px -2px;<br />
background-position: 0px -99px;<br />
}<br />
<br />
#mymenubar table div.biobrick1 {<br />
width: 70px;<br />
height: 95px;<br />
margin: 92px 20px 0px -2px ;<br />
background: url("https://static.igem.org/mediawiki/2013/c/c9/Biobrick26.png") no-repeat 0px 0px ;<br />
background-size: 70px 95px;<br />
}<br />
<br />
<br />
#mymenubar table td span.notebook {<br />
display: block;<br />
width: 70px;<br />
height: 95px;<br />
}<br />
<br />
#mymenubar table td.notebook {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 95px 20px 0px 0px;<br />
background: url("https://static.igem.org/mediawiki/2013/7/78/Notebook28.png") no-repeat 0px 0px;<br />
}<br />
<br />
#mymenubar table td.notebook:hover {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 95px 20px 0px 0px;<br />
background-position: 0px -100px;<br />
}<br />
<br />
#mymenubar table div.notebook1 {<br />
width: 70px;<br />
height: 95px;<br />
margin: 92px 20px 0px 0px ;<br />
background: url("https://static.igem.org/mediawiki/2013/8/8d/Notebook26.png") no-repeat 0px 0px ;<br />
background-size: 70px 95px;<br />
}<br />
<br />
#mymenubar table td span.humanpractice {<br />
display: block;<br />
width: 70px;<br />
height: 97px;<br />
}<br />
<br />
#mymenubar table td.humanpractice {<br />
float: left;<br />
width: 70px;<br />
height: 97px;<br />
margin: 86px 20px 0px -1px;<br />
background: url("https://static.igem.org/mediawiki/2013/c/c1/Humanpractice28.png") no-repeat 0px 0px;<br />
}<br />
<br />
#mymenubar table td.humanpractice:hover {<br />
float: left;<br />
width: 70px;<br />
height: 97px;<br />
margin: 86px 20px 0px -1px;<br />
background-position: 0px -99px;<br />
}<br />
<br />
#mymenubar table div.humanpractice1 {<br />
width: 70px;<br />
height: 95px;<br />
margin: 92px 20px 0px -1px ;<br />
background: url("https://static.igem.org/mediawiki/2013/5/54/Humanpractice26.png") no-repeat 0px 0px ;<br />
background-size: 70px 95px;<br />
}<br />
<br />
#mymenubar table td span.achievement {<br />
display: block;<br />
width: 70px;<br />
height: 95px;<br />
}<br />
<br />
#mymenubar table td.achievement {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 93px 20px 0px -2px;<br />
background: url("https://static.igem.org/mediawiki/2013/5/5e/Achievement28.png") no-repeat 0px 0px;<br />
}<br />
<br />
#mymenubar table td.achievement:hover {<br />
float: left;<br />
width: 70px;<br />
height: 95px;<br />
margin: 93px 20px 0px -2px;<br />
background-position: 0px -99px;<br />
}<br />
<br />
#mymenubar table div.achievement1 {<br />
width: 70px;<br />
height: 95px;<br />
margin: 92px 20px 0px -2px ;<br />
background: url("https://static.igem.org/mediawiki/2013/5/5a/Achievement26.png") no-repeat 0px 0px ;<br />
background-size: 70px 95px;<br />
}<br />
<br />
#index {<br />
float: left;<br />
width:490px;<br />
height:8000px;<br />
background: url("https://static.igem.org/mediawiki/2013/d/d6/Header_index_kiso5.png") 0px -250px no-repeat ;<br />
}<br />
<br />
<br />
#index img.logo {<br />
position: relative;<br />
top:20px;<br />
left:230px;<br />
width:150px;<br />
height:150px;<br />
}<br />
<br />
#index_title {<br />
position: relative;<br />
top:40px;<br />
left:230px;<br />
}<br />
<br />
#contents {<br />
position: relative;<br />
width:100px;<br />
top:240px;<br />
left:220px;<br />
}<br />
<br />
#contents2 {<br />
position: relative;<br />
width: 100px;<br />
top: 340px;<br />
left: 220px;<br />
}<br />
<br />
#main {<br />
float: left;<br />
width:820px;<br />
height:8000px;<br />
background: url("https://static.igem.org/mediawiki/2013/d/d6/Header_index_kiso5.png") -490px -250px no-repeat;<br />
<br />
}<br />
<br />
#space {<br />
float: left;<br />
width: 100px;<br />
height: 8000px;<br />
background: url("https://static.igem.org/mediawiki/2013/d/d6/Header_index_kiso5.png") -1310px -250px no-repeat;<br />
}<br />
<br />
#footerr {<br />
clear: both;<br />
width: 1410px;<br />
height: 700px;<br />
background: url("https://static.igem.org/mediawiki/2013/d/d6/Header_index_kiso5.png") 0px -9300px no-repeat;<br />
}<br />
<br />
#footer_sponsors ul {<br />
margin-left: 300px;<br />
}<br />
<br />
#footer_sponsors li {<br />
float: left;<br />
list-style-type: none;<br />
height: 70px;<br />
}<br />
<br />
#footer_sponsors li.UI {<br />
width:150px;<br />
margin: 50px 0px 0px 370px;<br />
}<br />
<br />
#footer_sponsors li.CB {<br />
width: ;<br />
margin: 80px 0px 0px 50px;<br />
}<br />
<br />
#footer_sponsors li.LN {<br />
width:;<br />
margin: 50px 0px 0px 50px;<br />
}<br />
<br />
#footer_sponsors li.IR {<br />
clear: both;<br />
width:;<br />
margin: 45px 0px 0px 370px;<br />
}<br />
<br />
#footer_sponsors li.Promega {<br />
width:;<br />
margin: 30px 0px 0px 50px;<br />
}<br />
<br />
#footer_sponsors li.IDT {<br />
width:;<br />
margin: 55px 0px 0px 50px;<br />
}<br />
<br />
#footer_sponsors li.MBL {<br />
width:;<br />
margin: 50px 0px 0px 370px;<br />
}<br />
<br />
#footer_sponsors li.NK {<br />
clear: both;<br />
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<h1 id="index_title"></h1><br />
<ul id="contents"><br />
<li><a href="#Introduction"><strong>Introduction</strong></a></li><br />
<li><a href="#Objective"><strong>Objective</strong></a></li><br />
<li><a href="#Method"><strong>Method</strong></a><br />
<ul><br />
<li><a href="#Design ">Design </a></li><br />
<li><a href="#Parts construction">Parts construction</a></li><br />
</ul><br />
</li> <br />
<li><a href="#Evaluation"><strong>Evaluation</strong></a><br />
<ul><br />
<li><a href="#TR-HHR">TR-HHR</a></li><br />
<li><a href="#TR-HHR-taRNA">TR-HHR-taRNA</a></li><br />
</ul><br />
</li><br />
<li><a href="#Future work"><strong>Future work</strong></a></li><br />
<br />
</ul><br />
<br />
<ul id="contents2"><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/oscillator"><li>RNA oscillator</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/scaffold"><li>RNAScaffold</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/light"><li>Light sensor</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/modeling"><li>Modeling</li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/rhodopsin"><li>Improving a BioBrick part - Rhodopsin</li></a><br />
</ul><br />
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<div id="main"><br />
<br />
<font size=5><br />
<BR><br />
<p align=center><font size=7><strong>RNA Oscillator</strong></font><hr></p><br />
<BR><br />
<BR><br />
<BR><br />
<font size=6><strong>Introduction </span></strong></font><hr><br />
<BR><br />
<BR><br />
<BR><br />
<font size=6 ><strong>Protein oscillator</span></strong></font><br />
<BR><br />
<BR><br />
<p style="line-height:110%"><br />
(There are many kinds of gene regulation system. One of them is repressor system that is used...)<br />
<BR><br />
The oscillating network was built by using three transcriptional repressor systems in which the first repressor protein suppresses the expression of second repressor gene, whose protein suppresses the transcription of the third gene, and the protein the third gene code suppresses the first gene expression.<br />
</p><br />
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<BR><br />
<BR><br />
<br />
<font size=6><strong>Hummer headed ribozyme</span></strong></font><br />
<br />
<BR><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
Hummerheaded ribozymes (HHRs) are found in many species and known to have a capability to self-cleave. Recently, engineered HHRs responding to trans-acting RNAs (taRNAs) in <I>E.coli</I> were reported. In absence of taRNA, the HHRs can self-cleave, but in presence of taRNA, their capability of self-cleavage is inhibited due to hybridize with taRNA and change their structures.<br />
</p><br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/3/3a/%E3%82%AD%E3%83%A3%E3%83%97%E3%83%81%E3%83%A318.PNG><BR><br />
Fig. 1 taRNAs responsible HHR<br />
<BR><BR><BR><br />
<br />
<p style="line-height:110%"><br />
We developed the new oscillator system that is composed only RNAs not using repressor gene and protein. We created three taRNA connected hummerheaded ribozymes (HHR-taRNA) that sense to another taRNA. The same way as the protein oscillator, three HHR-taRNAs inhibits other one’s self-cleavage such as three-way standoff. When the first HHR-taRNA is self-cleaved, connected the first taRNA that binds to the second HHR-taRNA is free and suppresses the second HHR-taRNA self-cleavage. When second HHR-taRNA is self-cleaved, the second taRNA that binds to the third HHR-taRNA is free and suppresses the third HHR-taRNA. Finally, when third HHR-taRNA is self-cleaved, the third taRNA that binds to the first HHR-taRNA is free and suppresses the first HHR-taRNA, completing cycle.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
<font size=6 id="Objective"><strong>Objective</span></strong></font><hr> <br />
<BR><br />
<BR><br />
Construction of Three of orthogonal taRNA generator which repressed by another taRNA.<br />
<br />
<BR><br />
<br />
<p style="line-height:110%"><br />
taRNA responsive HHR (TR-HHR) was fused with taRNA. By using orthogonal riboregulator sequences, we designed three of taRNA generators.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
<font size=6 id="Method"><strong>Method</span></strong></font><hr><br />
<br />
<BR><BR><br />
<br />
<font size=5 id="Design"><strong>Design</span></strong></font> <br />
<br />
<BR><BR><br />
<br />
<p style="line-height:110%"><br />
TR-HHR was designed by Benedikt Klauser and Jo¨ rg S. Hartig. Their TR-HHR were inspired by the riboregulatory system previously reported by Collins and coworkers. They made use of sequence domains of the riboregulator RR12. And, orthogonal variants of riboregulator RR12 (RR12y, RR42) were reported by Collins and coworkers. So, by using the sequences of these orthogonal riboregulators, we designed three orthogonal TR-HHRs.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
図1<br />
<br />
<BR><BR><BR><br />
<br />
<font size=5 id="Parts construction"><strong>Parts construction</span></strong></font> <br />
<BR><br />
<p style="line-height:110%"><br />
1. Construction of TR-HHR<BR><br />
2. Construction of TR-HHR variants<BR><br />
3. Construction of taRNA variants<BR><br />
4. Construction of TR-HHR-taRNA<BR><br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
1. Construction of TR-HHR<BR><br />
<br />
<p style="line-height:110%"> <br />
We used the TR-HHR developed by Klauser <I>et al.</I>[1] Two overlap primers designed, and PCR was performed to connect them. The PCR product was ligated into pSB1C3 vector.<br />
</p><br />
<br />
<BR><br />
<br />
<img src="https://static.igem.org/mediawiki/igem.org/e/e3/Wiki1.png" width="400px" / ><br />
<br />
<BR><BR><br />
<br />
2.Construction of TR-HHR variants<br />
<br />
<BR><BR><BR><br />
<br />
<p style="line-height:110%"><br />
By inverse PCR using pSB1C3-TR-HHR as a template, we could obtain three TR-HHRs, TR(12)-HHR, TR(12y)-HHR, and TR(42)-HHR.<br />
</p><br />
<br />
<img src="https://static.igem.org/mediawiki/igem.org/9/98/Wiki6.png" width="400px" / ><br />
<br />
<BR><br />
<BR><br />
<BR><br />
<br />
<br />
3.Construction of taRNA variants<br />
<br />
<BR><br />
<BR><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
By inverse PCR using pSB1A3-taR12-DT as a template, we could obtain three taRNAs, taR12, taR12y, taR42.<br />
</p><br />
<br />
<img src="https://static.igem.org/mediawiki/igem.org/a/a7/Wiki2.png" width="400px" / ><br />
<br />
<BR><br />
<BR><br />
<BR><br />
<br />
4.Construction of TR-HHR-taRNA<br />
<BR><br />
<BR><br />
<BR><br />
<br />
TR-HHR-taRNAs were constructed by overlap PCR.<br />
<br />
<BR><br />
<BR><br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/9/91/Wiki3.png" width="400px" / ><br />
<br />
<BR><br />
<BR><br />
<BR><br />
<BR><br />
<BR><br />
<BR><br />
<font size=6 id="Evaluation"><strong>Evaluation</strong></font><hr><br />
<BR><br />
<BR><br />
I. Evaluation of TR-HHR<br />
<BR><BR><br />
II. Evaluation of TR-HHR-taRNA<br />
<br />
<BR><BR><BR><BR><br />
<br />
<font size=6 id="TR-HHR"><strong>I. Evaluation of TR-HHR</strong></font><br />
<br />
<BR><br />
<br />
<p style="line-height:110%"><br />
In order to evaluate the regulation of HHR’s self-cleavage activity by taRNA, GFP gene was connected with the HHR or inactive HHR (*HHR), which has a mutation lose the activity. These fragments were subcloned into conventional biobrick standard vector pSB3C5. <br />
</p><br />
<br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/4/45/Wiki4.png" width="400px" / ><br />
<BR><br />
<br />
1) <I>E. coli</I> TOP10 strain was co-transformed by two plasmids listed bellow, <br />
<BR><br />
<BR><br />
A)<br />
<BR><br />
pSB1A3-taR12-DT<br />
<BR><br />
pSB3C5-TR(12)-HHR -GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
B)<br />
<BR><br />
pSB1A3-empty<br />
<BR><br />
pSB3C5- TR(12)-HHR -GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
<br />
C) <br />
<BR><br />
pSB1A3-taR12-DT<br />
<BR><br />
pSB3C5-TR(12)- * HHR-GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
<br />
D)<br />
<BR><br />
pSB1A3 empty<br />
<BR><br />
pSB3C5- TR(12)- * HHR -GFP-DT<br />
<br />
<BR><br />
<BR><br />
<br />
<br />
E) <br />
<BR>pSB1A3-empty<br />
<BR><br />
pSB3C5-empty<br />
<br />
<BR><br />
<BR><br />
<br />
<p style="line-height:110%"><br />
2) Bacterial cultures were incubated in LB medium at 140 rpm and 37℃ for 12 hour. <BR><br />
3) Then, cultures was diluted to an OD595 of 0.0125 and induced with or without 0.1% arabinose.<BR><br />
4) Cultures were incubated with shaking at 140 rpm and 37℃ for 8 hours in 1 mL of LB medium.<BR><br />
5) OD595 and GFP fluorescence intensity were measured.<BR><br />
All experiments were performed using three cultures per each sample.<br />
</p><br />
<br />
<BR><BR><BR><br />
<br />
Result and Discussion<br />
<br />
<BR><BR><br />
<br />
<p style="line-height:110%"><br />
In the presence of taR12, GFP fluorescence value divided by OD value is higher than one in absence of taR12. This result was not expected. HHR has the ability to self-cleave, but its ability is suppressed when taRNA hybridizes to HHR. Therefore, in the presence of taRNA, self-cleavage of HHR should not happen. So then, why we get such result? One factor can be due to the exposure of RBS sequence caused by the structural change of HHR when it hybridizes to taRNA<br />
</p><br />
<br />
<BR><BR><BR><BR><br />
<br />
<font size=6 id="TR-HHR-taRNA">II.Evaluation of TR-HHR-taRNA</font><br />
<br />
<BR><BR><br />
<br />
We examined whether TR-HHR-taRNA’s output can be repressed by corresponding taRNA. <br />
<BR><br />
<BR><br />
<img src="https://static.igem.org/mediawiki/igem.org/5/52/Wiki5.png" width="400px" / ><br />
<BR><br />
<br />
<br />
1) The plasmids as shown below was used to transform <I>E. coli </I>Top10.<BR><br />
<BR><br />
A) <br />
<BR><br />
pSB1A3-P<sub>bad</sub> -TR(42)HHR-taR12-DT<br />
<BR><br />
pSB3C5-crR12-GFP<br />
<br />
<BR><BR><BR><br />
<br />
B) <br />
<BR><br />
pSB1A3-P<sub>const(H)</sub>-taR42-P<sub>bad</sub>-TR(42)HHR-taR12-DT<BR><br />
pSB3C5-crR12-GFP-DT<br />
<br />
<BR><BR><BR><br />
<br />
C)<br />
<BR><br />
pSB1A3-P<sub>bad</sub>-taR12-DT<BR><br />
pSB3C5-crR12-GFP<br />
<br />
<BR><BR><BR><br />
<br />
D) <BR><br />
pSB1A3-empty <BR><br />
pSB3C5-crR12-GFP<br />
<br />
<BR><BR><BR><br />
<br />
E) <BR><br />
pSB1A3-empty<BR><br />
pSB3C5-empty<BR><br />
<br />
<BR><BR><br />
<br />
<p style="line-height:110%"><br />
2) The transformants were pre-cultured in LB medium at 37 degrees celcius for 8 hours. <BR><br />
3) Then, cultures was diluted to an OD595 of 0.0125 and induced with or without 0.1% arabinose.<BR><br />
4) Cultures were incubated with shaking at 140 rpm and 37℃ for 8 hours in 1 mL of LB medium.<BR><br />
5) OD595 and GFP fluorescence intensity were measured.<BR><br />
</p><br />
<br />
<BR><BR><br />
<br />
<font size=5 ><strong>Result and Discussion</strong></font><hr><br />
<br />
<BR><br />
<BR><br />
<p style="line-height:110%"><br />
In order to evaluate a function of TR(42)-HHR-taR12, we used crR12-GFP as a reporter.<br />
taRNA is complementary to the sequence of crRNA, and by the binding of taRNA to crRNA, RBS is exposed and gene expression is activated. In this result, the GFP fluorescence value divided by OD value of (2) was lower than that of (3). It shows that taR42 suppressed self-cleavage of TR(42)-HHR and caused less GFP expression. Therefore, we prove that the self-cleavage activity of TR(42)-HHR and amount of taR12 can be controlled by taR42.<br />
</p><br />
<br />
<BR><BR><BR><BR><br />
<br />
<font size=6 id="Future work" ><strong>・Future work</strong></font><hr><br />
<br />
<BR><BR><BR><BR><BR><br />
<br />
<font size=6 ><strong>・Reference</strong></font><br />
<br />
<BR><BR><br />
<BR><br />
<p style="line-height:110%"><br />
[1]<br />
An engineered small RNA-mediated genetic switch<br />
based on a ribozyme expression platform<br />
Benedikt Klauser and Jo¨ rg S. Hartig <br />
Nucleic Acids Research, 2013<br />
<BR><br />
[2]<br />
Callura,J.M., Cantor,C.R. and Collins,J.J. (2012) Genetic<br />
switchboard for synthetic biology applications. Proc. Natl Acad.<br />
Sci. USA, 109, 5850–5855<br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Daily_notes
Team:Sumbawagen/Daily notes
2013-09-28T02:41:12Z
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Lab_Preparation_team_activities
Team:Sumbawagen/Lab Preparation team activities
2013-09-28T02:40:13Z
<p>Jinhee: </p>
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Lab_Preparation_team_activities
Team:Sumbawagen/Lab Preparation team activities
2013-09-28T02:39:35Z
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Jinhee
http://2013.igem.org/File:%E3%82%B9%E3%82%AF%E3%83%AA%E3%83%BC%E3%83%B3%E3%82%B7%E3%83%A7%E3%83%83%E3%83%88_2013-09-28_11.21.47.png
File:スクリーンショット 2013-09-28 11.21.47.png
2013-09-28T02:25:31Z
<p>Jinhee: </p>
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Lab_Preparation_team_activities
Team:Sumbawagen/Lab Preparation team activities
2013-09-28T02:20:07Z
<p>Jinhee: </p>
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Jinhee
http://2013.igem.org/File:L6.png
File:L6.png
2013-09-28T02:19:32Z
<p>Jinhee: </p>
<hr />
<div></div>
Jinhee
http://2013.igem.org/File:L5.png
File:L5.png
2013-09-28T02:16:03Z
<p>Jinhee: </p>
<hr />
<div></div>
Jinhee
http://2013.igem.org/Team:Sumbawagen/Lab_Preparation_team_activities
Team:Sumbawagen/Lab Preparation team activities
2013-09-28T02:13:53Z
<p>Jinhee: </p>
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Acknowledgement
Team:Sumbawagen/Acknowledgement
2013-09-28T02:13:07Z
<p>Jinhee: </p>
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<p style="padding-top: 0pt; " class="Free_Form">We would like to extend gratitude for supporting our team in iGEM project to :<br /></p><br />
<p class="Free_Form"><br /></p><br />
<p class="Free_Form">1.Dr. Zulkifliemansyah, M. Sc (Rector of Sumbawa University of Technology)<br /></p><br />
<p class="Free_Form">For the opportunity, supports, patient and trust to our team<br /></p><br />
<p class="Free_Form"><br /></p><br />
<p class="Free_Form">2.Dr. Arief Budi Witarto, Ph. D (Dean of Technobiology Faculty, SUT)<br /></p><br />
<p class="Free_Form">For the opportunity and trust <br /></p><br />
<p class="Free_Form"><br /></p><br />
<p class="Free_Form">3.Prof. Koji Sode, PhD<br /></p><br />
<p class="Free_Form">For visiting our team and gave precious suggestions to improve our skill and motivation.<br /></p><br />
<p class="Free_Form"><br /></p><br />
<p class="Free_Form">4.Dr. Tomohiko Yamazaki<br /></p><br />
<p class="Free_Form">For visiting our team and sharing experience about research skill.<br /></p><br />
<p class="Free_Form"><br /></p><br />
<p class="Free_Form">5.Mr. Bugi Ratno Budiharjo, S.Si, M.Sc<br /></p><br />
<p class="Free_Form">For your participation during ‘Biotech Camp’ and ‘Biotechnology Competition 2’ in Sumbawa University of Technology.<br /></p><br />
<p class="Free_Form"><br /></p><br />
<p class="Free_Form">6.Mr. Aka Kurnia SF, S.Ag<br /></p><br />
<p class="Free_Form">For taking nice pictures of our team.<br /></p><br />
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<p class="Free_Form">7.Tokyo-NoKoGen<br /></p><br />
<p class="Free_Form">For making our team wiki.<br /></p><br />
<p class="Free_Form"><br /></p><br />
<p class="Free_Form">8.Ms. Dwi Ariyanti, S.Pt, M.Biotech and Ms. Maya Fitriana, S.Si<br /></p><br />
<p class="Free_Form">Our new instructors, for helping us finishing our project.<br /></p><br />
<p class="Free_Form"><br /></p><br />
<p class="Free_Form">9.Government, students, and society of Sumbawa regency and West Sumbawa regency<br /></p><br />
<p class="Free_Form">For the participation in the survey for human practice project.<br /></p><br />
<p class="Free_Form"><br /></p><br />
<p class="Free_Form">10.All staffs of Sumbawa University of Technology<br /></p><br />
<p style="padding-bottom: 0pt; " class="Free_Form">For supporting us to finish our project.</p><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Lab_Preparation_team_activities
Team:Sumbawagen/Lab Preparation team activities
2013-09-28T02:11:29Z
<p>Jinhee: </p>
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Jinhee
http://2013.igem.org/File:L4.png
File:L4.png
2013-09-28T02:10:54Z
<p>Jinhee: </p>
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Lab_Preparation_team_activities
Team:Sumbawagen/Lab Preparation team activities
2013-09-28T02:02:39Z
<p>Jinhee: </p>
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Jinhee
http://2013.igem.org/File:L3.png
File:L3.png
2013-09-28T02:00:51Z
<p>Jinhee: </p>
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<div></div>
Jinhee
http://2013.igem.org/File:L2.png
File:L2.png
2013-09-28T01:49:03Z
<p>Jinhee: </p>
<hr />
<div></div>
Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/questionnaire
Team:Tokyo-NoKoGen/questionnaire
2013-09-27T20:36:35Z
<p>Jinhee: </p>
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<table><br />
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<br />
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<br />
<br />
<h1 id="index_title"></h1><br />
<ul id="contents"><br />
<a href="#overview"><li><strong>What is synthetic biology?</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/labosafety"><li><strong>Safety in the laboratoy</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/questionnaire"><li><strong>Questionnaire on synthetic biology</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/ikalux"><li><strong>Do It Yourself -Ika Lux-</strong></li></a><br />
<br />
</ul><br />
<br />
</div><br />
<br />
<div id="main"><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=6>Questionnaire on synthetic biology</font></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>We would like to thank all the people who helped us answer the questionnaire, we were able to get 85 replies by 2 p.m. on 27th of September. We prepared 21 questions to find out how people recognize synthetic biology. </p></font><br />
<br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q1. Are you a male or a female?</font></p><br />
<p align=center><font size=3>性別を教えてください</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/5/52/%E3%82%B9%E3%83%A9%E3%82%A4%E3%83%891.JPG></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>The ratio is almost ideal!</font></p><br />
<br />
<BR><br />
<p align=center><font size=4>Q2. What age are you?</font></p><br />
<p align=center><font size=3>年代を教えてください</p></font><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/1/1f/Q2.JPG></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Unfortunately, the ratio of generations is biased. This means that this questionnaire tend to reflect younger people's opinion.</font></p><br />
<BR><br />
<p align=center><font size=4>Q3. Are you specialized in science or non-science?</font></p><br />
<p align=center><font size=3>文系か理系、どちらがお得意かお答えください</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/2/2c/Q3.JPG></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Most of people are major in science. </font></p><br />
<BR><br />
<p align=center><font size=4>Q4. Do you know synthetic biology?</font></p><br />
<p align=center><font size=3>「合成生物学」を知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/5/52/Q4.JPG></p><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Even though many people who involved in this questionnaire are major in science, it seems that the word "Synthetic biology" is unfamiliar.</font></p><br />
<BR><br />
<p align=center><font size=4>Q5. If yes, how did you get to know about it?</font></p><br />
<p align=center><font size=3> 「はい」と答えた方はどこで知りましたか?<br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/8/80/Q5.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q6. What do you think Synthetic biology is about?</font></p><br />
<p align=center><font size=3> 合成生物学は次のうちどのような学問だと思いますか?<br />
<p alignt=center><img src=https://static.igem.org/mediawiki/2013/f/f9/Q6.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q7. Do you know iGEM?</font></p><br />
<p align=center><font size=3>iGEMという大会を知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/1/11/Q7.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q8. If yes, how do you know it?</font></p><br />
<p align=center><font size=3>「はい」と答えた方はどこで知りましたか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/9/9f/Q8.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q9. How do you pronounce "iGEM"?</font></p><br />
<p align=center><font size=3>"iGEM"の読み方は次のうちどれだと思いますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/0/04/Q9.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>10. Where do you think the iGEM Jamboree is held?</font></p><br />
<p align=center><font size=3>iGEMの本選はどこで開催されていると思いますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/1/19/Q10.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q11. What do you think is important in iGEM?</font></p><br />
<p align=center><font size=3>iGEMとはどのようなことを競う大会だと思いますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/4/4b/Q11.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q12. How many Japanese teams do you think participate iGEM?</font></p><br />
<p align=center><font size=3>iGEMの日本のチームは何チームいるか知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/e/e1/Q12.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q13. How many teams do you think there are in the whole iGEM society?</font></p><br />
<p align=center><font size=3>世界にはiGEMが何チームいるか知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/8/8f/Q13.JPG></P><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q14. What do you think of, when you hear "E. coli"?</font></p><br />
<p align=center><font size=3>「大腸菌」という単語から連想するものは何ですか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/c/cf/Q14.JPG></P><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q15. What do you think of, when you hear "Genetic modification"?</font></p><br />
<p align=center><font size=3>「遺伝子組み換え」という単語から連想するものは何ですか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/4/4b/Q15.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q16. Did you know that genetically modified E. coli can collect heavy metal ions?</font></p><br />
<p align=center><font size=3>遺伝子組み換え型大腸菌を使って重金属を回収することが出来ることを知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/a/a0/Q16.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q17. Did you know that genetically modified E. coli can become luminescent?</font></p><br />
<p align=center><font size=3>遺伝子組み換え型大腸菌が青色に光ることが出来ることを知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/4/40/Q17.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q18. Did you know that genetically modified E. coli can be regulated by light?</font></p><br />
<p align=center><font size=3>遺伝子組み換え型大腸菌を光で制御出来ることを知っていますか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/2/22/Q18.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q19. We bought a squid at a nearby supermarket. What do you think we did with them?</font></p><br />
<p align=center><font size=3>私達は近所のスーパーで石川県産のイカを買いました。?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/e/e6/Q19.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q20. After this questionnaire, do you think synthetic biology is interesting?</font></p><br />
<p align=center><font size=3>合成生物学について、なんとなく興味を持っていただけたでしょうか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/d/db/Q20.JPG></p><br />
<BR><br />
<BR><br />
<BR><br />
<p align=center><font size=4>Q21. Do you also feel that you want to participate iGEM competitiong?</font></p><br />
<p align=center><font size=3>iGEMに参加したいと思ってもらえたでしょうか?</font></p><br />
<p align=center><img src=https://static.igem.org/mediawiki/2013/5/5a/Q21.JPG></p><br />
<BR><br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen
Team:Tokyo-NoKoGen
2013-09-27T20:19:40Z
<p>Jinhee: </p>
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<h1 id="index_title"></h1><br />
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<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/oscillator"><li><a href="#diary"><strong>RNA oscillator</a></strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/scaffold"><li><strong>RNA scaffold</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/light"><li></strong>Light sensor</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/modeling"><li><strong>Modeling</strong></li></a><br />
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<p align=center><font size=7>Introduction</font></p></font><br />
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<p style="text-indent:2em"><font size=5><br />
We all have a biological clock which controls the periodicity of<br />
many physiological functions such as blood pressure, body temperature and concentration of hormones. The systems that generate circadian rhythms are called oscillator. Oscillator has been researched for deepening our knowledge of circadian rhythms,understanding genetic network and signal transfer.</p><br />
<BR><br />
<div style="text-align:center"><br />
<img src="https://static.igem.org/mediawiki/2013/0/0d/Circadian_rhythm.PNG", height="300", width="450",alt=""><br />
</div><br />
<div style="text-align:center"><br />
Fig.1 Circadian rhythm<br />
</div><br />
<p style="text-indent:2em"><br />
Oscillator is expected to be used for various applications.</p><br />
<BR><br />
<p style="text-indent:2em"><br />
Therefore, desired features of oscillator are:</p><br />
<p style="text-indent:4em"><br />
1. Variation in types</p><br />
<p style="text-indent:4em"><br />
2. Feasibility in designing and controlling</p><br />
<BR><br />
<p style="text-indent:2em"><br />
However, existing protein oscillator does not satisfy the above requirements.</p><br />
<img src="https://static.igem.org/mediawiki/2013/9/91/Protein.PNG",height="200", width="200"alt="" style="float:right"><br />
<p style="text-indent:2em"><br />
The first problem of protein oscillator is the small type of the protein. Because the number of repressor proteins and promoters are limited, the variation of protein oscillator is not many. </p><br />
<p style="text-indent:2em"><br />
Another problem of protein oscillator is the component, the protein. Protein requires several steps; transcription and translation, protein modification, folding, and degradation. When we want to construct or control such protein oscillator, we have to genetically engineer to change the transcriptional efficiency, translational efficiency and degradation ability. Therefore, it is difficult to design such gene circuits that can express the repressors periodically. </p><br />
<BR><br />
<BR><br />
<BR><br />
<p style="text-indent:2em"><br />
To satisfy the requirements, we propose the New type of oscillator circuit – RNA oscillator.</p><br />
<br />
<p style="text-indent:2em"><br />
Whereas there are only a few types of proteins that can be used for construction of protein oscillator, many kinds of RNA oscillators can be constructed by changing a few base pair. Therefore, it will allow us to construct a multiple and orthogonal oscillators.</p><br />
<p style="text-indent:2em"><br />
Another advantage is that, although protein has several steps until degradation, RNA has no steps of translation, modification and time-consuming degradation. Therefore, RNA oscillator can be designed and controlled by only changing the transcriptional efficiency and binding ability. It is easy to change the binding ability of RNA because all we have to do is to alter the base pair, and hence change the strength of base pairs binding.</p><br />
<br />
<img src="https://static.igem.org/mediawiki/2013/5/57/ProteinRNA.PNG",height="300",width="300",alt=""><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/Alter.PNG", height="300", width="300", alt="",style="float:right"><br />
<p style="text-indent:2em"><br />
Like this, RNA meets the necessary standards for:</p><br />
<p style="text-indent:4em"><br />
1. Variation in types</p><br />
<p style="text-indent:4em"><br />
2. Feasibility in designing and controlling</p><br />
<BR><br />
<p style="text-indent:2em"><br />
Therefore, it can be used for combination with diverse mechanisms.<br />
For example, we can change output signals from RNA to other ones such as protein with reporter circuit, and control the oscillation with light sensor.</p><br />
<br />
<p style="text-indent:2em"><br />
In this way, the ability to combine with various mechanisms can be used for controlling complex systems such as drug delivery.</p><br />
<p style="text-indent:2em"><br />
We named the E. coli which has this function TwinklE.coli because of its brilliant future.<br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement
Team:Tokyo-NoKoGen/Acknowledgement
2013-09-27T19:56:20Z
<p>Jinhee: </p>
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<p align=center><font size=7>Acknowledgement</font></p><br />
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<BR><font size=4><br />
<br />
<br />
We would like to extend gratitude for supporting our team in iGEM project to :<br />
<BR><BR><br />
1. Prof. Koji Sode<BR><br />
For the opportunity, supports, patient and trust to our team<br />
<BR><BR><br />
2. Prof. Kazunori Ikebukuro<BR><br />
For the opportunity and trust <br />
<BR><BR><br />
3. Prof. Stefano Ferri<BR><br />
For the opportunity and trust, giving advise for experiments, and improving our English skill.<br />
<BR><BR><br />
4. Prof. Wakako tusgawa<BR><br />
For the opportunity and trust <br />
<BR><BR><br />
5. Dr. Koichi Abe<BR><br />
For helping us finishing our project.<br />
<BR><BR><br />
6. iGEM Japan<BR><br />
For sharing ideas and information<br />
<BR><BR><br />
7. All staffs of Tokyo University of Agriculture and Technology<BR><br />
For supporting us to finish our project.<br />
<BR><BR><br />
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Jinhee
http://2013.igem.org/File:L1.png
File:L1.png
2013-09-27T19:16:17Z
<p>Jinhee: </p>
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Lab_Preparation_team_activities
Team:Sumbawagen/Lab Preparation team activities
2013-09-27T19:15:09Z
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Team
Team:Sumbawagen/Team
2013-09-27T18:54:16Z
<p>Jinhee: </p>
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<h2>Advisor</h2><br />
<br><br />
Dr. Zulkieflimansyah <br />
<br><br />
<h2>instructor</h2><br />
<br><br />
Dr. Arief Budi Witarto<br />
<br><br />
Julmansyah<br />
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<h2>Student Member</h2><br />
<br><br />
Fahmi Dwilaksono<br />
<br><br />
Abdhi Surya Perkasa<br />
<br><br />
Adelia Elviantari<br />
<br><br />
Arif Muhammad<br />
<br><br />
Asmawati<br />
<br><br />
Cendra Harri Pahlawan<br />
<br><br />
Cindy Suci Ananda<br />
<br><br />
Gita Feny Lestari<br />
<br><br />
Indah Nurulita<br />
<br><br />
M. Al Azhar<br />
<br><br />
M. Isro Al Fajri<br />
<br><br />
Nugrah Analiadi Putra<br />
<br><br />
Rian Adha Ardinata<br />
<br><br />
Yulianti<br />
<br><br />
Yunianti<br />
<br><br />
<br><br />
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<h2> </h2><br />
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Fahmi Dwilaksono<br />
<br><br />
<br />
<IMG SRC="https://static.igem.org/mediawiki/2013/thumb/0/0c/Fahmi.png/469px-Fahmi.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Fahmi Dwilaksono as Leader was born April, 19th 1995,Grade 1st, Joinning iGEM is a big prize for me. I know I have to work hard and study hard everyday. But I have a solid team : my lovely advisors, instructors, and of course my teammates, so I enjoy this project. I learn a lot about synthetic biology, and I realize it’s very useful for life. It would be excited to see other teams around the world, and hopefully we will get the best achievement in HongKong. Fighting!!!<br />
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<br><br />
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Abdi surya perkasa<br />
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<IMG SRC="https://static.igem.org/mediawiki/2013/4/4d/Abdi.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Abdi surya perkasa was born July, 14th 1996,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best<br />
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<br><br />
<br><br />
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<br />
Adelia Elviantari<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/b/bb/433px-Adel.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Adelia Elviantari was born June,26th 2013,Grade 1st,I am grateful to be part of from sumbawagen iGEM team 2013. Because I get the experience that I had never experienced before during the iGEM 2013, me and my friends taught new things that open our eyes to the sophistication of the technology.<br />
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<br><br />
<br><br />
<br><br />
<br />
Arif Muhamad<br />
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<IMG SRC="https://static.igem.org/mediawiki/2013/8/86/Arif.png" WIDTH=120 HEIGHT=120 ><br />
<br><br />
<br />
Arif Muhamad was born March, 3rd 1995,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best.<br />
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<br><br />
<br />
Asmawati<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/3/3a/475px-Asmawati.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Asmawati was born May,8th 1995, Grade 1 st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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<br><br />
<br><br />
<br><br />
<br />
Cendra Harri Pahlawan<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/thumb/2/26/Cendra.png/180px-Cendra.png" WIDTH=100 HEIGHT=130 ><br />
<br><br />
<br />
<br />
Cendra Hari pahlawan was born November,10th 1994, Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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<br><br />
<br><br />
<br><br />
<br />
Cindy Suci Ananda<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/7/72/535px-Cindy.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Cindy Suci Ananda was born September, 16th 1996, Grade 1st, get a chance to join in and become part of the Sumbawagen is a boon for me, and I am very grateful with this grace, iGEM and sumbawagen taught the meaning of many things. Thick and thin, laughter and jokes, teamwork, es tired for lack of time to work but I am still grateful. Fighting friendships dissolve and become a family. Although sometime always Sumbawagen.<br />
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<br><br />
<br><br />
<br><br />
<br />
Gita Feny Lestari<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/f/fc/481px-Gita.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
<br />
Gita Feny lestari was born February,22th 1995, Grade 1st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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<br><br />
<br><br />
<br><br />
<br />
<br />
Indah Nurulita<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/9/98/Indah.png" WIDTH=100 HEIGHT=130 ><br />
<br><br />
Indah Nurulita was born March,28th 1995, Grade 1st, sumbawagen is a second home to me recently that I got a lot of experience with them, spend time with them gives a distinct impression. Although the task after task, but all come with their mild compactness feels real. spirit sumbawagen.<br />
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<br><br />
<br><br />
<br><br />
<br />
<br />
M. Al Azhar<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/2/2d/465px-Azhar.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Muhammad Al-Azhar was born june,25th 1995, Grade 1st, to be cool do not need an expert to play music, basketball or the other. enough to understand that science is cool from everything, and IGEM is a cool competition that must be followed.<br />
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<br><br />
<br><br />
<br><br />
<br />
<br />
M. Isro Al Fajri<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/a/a1/443px-Isro.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Mochammad isro alfajri was born December, 20th 1995, Grade 1st, and i especialist and leader in webdesign, humans and nature is like the driver with the vehicle. men that will bring natural conditions to welfare or destruction. because man is the leader in this world, and iGEM provides a view of nature to make it more meaningful.<br />
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<br><br />
<br><br />
<br><br />
<br />
Nugrah Analiadi Putra<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/6/6a/463px-Nugrah.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Nugrah Analiadi Putrawas born December,8th 1994, Grade 1st Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
---- <br />
<br><br />
<br><br />
<br><br />
<br />
Rian Adha Ardinata<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/8/85/428px-Rian.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Rian Adha ardinata was born May,9th 1995, Grade 1st, IGEM gives a new view to me. that life is a reciprocal relationship, whereby we must be able to utilize the natural resources to make this earth a smile.<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
Yulianti<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/b/b1/Yuli.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Yulianti was born july 1995,14th Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
Yunianti<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/f/f1/Yuni.png" WIDTH=110 HEIGHT=120 ><br />
<br><br />
<br />
Yunianti was born May,27th 1994, Grade 1st, IGEM is make my knowledge increase. IGEM makes me can to look nem other world never I look and feel.<br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Team
Team:Sumbawagen/Team
2013-09-27T18:53:34Z
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<p style="padding-bottom: 0pt; padding-top: 0pt; " class="Title">SUMBAWA GEN team member</p><br />
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<h2>Advisor</h2><br />
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Dr. Zulkieflimansyah <br />
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<h2>instructor</h2><br />
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Dr. Arief Budi Witarto<br />
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Julmansyah<br />
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<h2>Student Member</h2><br />
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Fahmi Dwilaksono<br />
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Abdhi Surya Perkasa<br />
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Adelia Elviantari<br />
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Arif Muhammad<br />
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Asmawati<br />
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Cendra Harri Pahlawan<br />
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Cindy Suci Ananda<br />
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Gita Feny Lestari<br />
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Indah Nurulita<br />
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M. Al Azhar<br />
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M. Isro Al Fajri<br />
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Nugrah Analiadi Putra<br />
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Rian Adha Ardinata<br />
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Yulianti<br />
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Yunianti<br />
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<h2> </h2><br />
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Fahmi Dwilaksono<br />
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Fahmi Dwilaksono as Leader was born April, 19th 1995,Grade 1st, Joinning iGEM is a big prize for me. I know I have to work hard and study hard everyday. But I have a solid team : my lovely advisors, instructors, and of course my teammates, so I enjoy this project. I learn a lot about synthetic biology, and I realize it’s very useful for life. It would be excited to see other teams around the world, and hopefully we will get the best achievement in HongKong. Fighting!!!<br />
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Abdi surya perkasa<br />
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Abdi surya perkasa was born July, 14th 1996,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best<br />
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Adelia Elviantari<br />
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Adelia Elviantari was born June,26th 2013,Grade 1st,I am grateful to be part of from sumbawagen iGEM team 2013. Because I get the experience that I had never experienced before during the iGEM 2013, me and my friends taught new things that open our eyes to the sophistication of the technology.<br />
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Arif Muhamad<br />
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Arif Muhamad was born March, 3rd 1995,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best.<br />
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Asmawati<br />
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Asmawati was born May,8th 1995, Grade 1 st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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Cendra Harri Pahlawan<br />
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Cendra Hari pahlawan was born November,10th 1994, Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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Cindy Suci Ananda<br />
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Cindy Suci Ananda was born September, 16th 1996, Grade 1st, get a chance to join in and become part of the Sumbawagen is a boon for me, and I am very grateful with this grace, iGEM and sumbawagen taught the meaning of many things. Thick and thin, laughter and jokes, teamwork, es tired for lack of time to work but I am still grateful. Fighting friendships dissolve and become a family. Although sometime always Sumbawagen.<br />
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Gita Feny Lestari<br />
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Gita Feny lestari was born February,22th 1995, Grade 1st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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Indah Nurulita<br />
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Indah Nurulita was born March,28th 1995, Grade 1st, sumbawagen is a second home to me recently that I got a lot of experience with them, spend time with them gives a distinct impression. Although the task after task, but all come with their mild compactness feels real. spirit sumbawagen.<br />
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M. Al Azhar<br />
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Muhammad Al-Azhar was born june,25th 1995, Grade 1st, to be cool do not need an expert to play music, basketball or the other. enough to understand that science is cool from everything, and IGEM is a cool competition that must be followed.<br />
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M. Isro Al Fajri<br />
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Mochammad isro alfajri was born December, 20th 1995, Grade 1st, and i especialist and leader in webdesign, humans and nature is like the driver with the vehicle. men that will bring natural conditions to welfare or destruction. because man is the leader in this world, and iGEM provides a view of nature to make it more meaningful.<br />
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Nugrah Analiadi Putra<br />
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Nugrah Analiadi Putrawas born December,8th 1994, Grade 1st Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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Rian Adha Ardinata<br />
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Rian Adha ardinata was born May,9th 1995, Grade 1st, IGEM gives a new view to me. that life is a reciprocal relationship, whereby we must be able to utilize the natural resources to make this earth a smile.<br />
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Yulianti<br />
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<IMG SRC="https://static.igem.org/mediawiki/2013/b/b1/Yuli.png" WIDTH=100 HEIGHT=120 ><br />
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Yulianti was born july 1995,14th Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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Yunianti<br />
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Yunianti was born May,27th 1994, Grade 1st, IGEM is make my knowledge increase. IGEM makes me can to look nem other world never I look and feel.<br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Team
Team:Sumbawagen/Team
2013-09-27T18:53:16Z
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<h2>Advisor</h2><br />
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Dr. Zulkieflimansyah <br />
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<h2>instructor</h2><br />
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Dr. Arief Budi Witarto<br />
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Julmansyah<br />
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<h2>Student Member</h2><br />
<br><br />
Fahmi Dwilaksono<br />
<br><br />
Abdhi Surya Perkasa<br />
<br><br />
Adelia Elviantari<br />
<br><br />
Arif Muhammad<br />
<br><br />
Asmawati<br />
<br><br />
Cendra Harri Pahlawan<br />
<br><br />
Cindy Suci Ananda<br />
<br><br />
Gita Feny Lestari<br />
<br><br />
Indah Nurulita<br />
<br><br />
M. Al Azhar<br />
<br><br />
M. Isro Al Fajri<br />
<br><br />
Nugrah Analiadi Putra<br />
<br><br />
Rian Adha Ardinata<br />
<br><br />
Yulianti<br />
<br><br />
Yunianti<br />
<br><br />
<br><br />
<br />
<h2> </h2><br />
<br />
Fahmi Dwilaksono<br />
<br><br />
<br />
<IMG SRC="https://static.igem.org/mediawiki/2013/thumb/0/0c/Fahmi.png/469px-Fahmi.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Fahmi Dwilaksono as Leader was born April, 19th 1995,Grade 1st, Joinning iGEM is a big prize for me. I know I have to work hard and study hard everyday. But I have a solid team : my lovely advisors, instructors, and of course my teammates, so I enjoy this project. I learn a lot about synthetic biology, and I realize it’s very useful for life. It would be excited to see other teams around the world, and hopefully we will get the best achievement in HongKong. Fighting!!!<br />
----<br />
<br><br />
<br><br />
<br><br />
Abdi surya perkasa<br />
<br><br />
<br />
<IMG SRC="https://static.igem.org/mediawiki/2013/4/4d/Abdi.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Abdi surya perkasa was born July, 14th 1996,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best<br />
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<br><br />
<br><br />
<br><br />
<br />
Adelia Elviantari<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/b/bb/433px-Adel.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Adelia Elviantari was born June,26th 2013,Grade 1st,I am grateful to be part of from sumbawagen iGEM team 2013. Because I get the experience that I had never experienced before during the iGEM 2013, me and my friends taught new things that open our eyes to the sophistication of the technology.<br />
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<br><br />
<br><br />
<br><br />
<br />
Arif Muhamad<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/8/86/Arif.png" WIDTH=120 HEIGHT=120 ><br />
<br><br />
<br />
Arif Muhamad was born March, 3rd 1995,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best.<br />
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<br><br />
<br />
Asmawati<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/3/3a/475px-Asmawati.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Asmawati was born May,8th 1995, Grade 1 st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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<br><br />
<br><br />
<br><br />
<br />
Cendra Harri Pahlawan<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/thumb/2/26/Cendra.png/180px-Cendra.png" WIDTH=100 HEIGHT=130 ><br />
<br><br />
<br><br />
<br><br />
<br />
Cendra Hari pahlawan was born November,10th 1994, Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
Cindy Suci Ananda<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/7/72/535px-Cindy.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Cindy Suci Ananda was born September, 16th 1996, Grade 1st, get a chance to join in and become part of the Sumbawagen is a boon for me, and I am very grateful with this grace, iGEM and sumbawagen taught the meaning of many things. Thick and thin, laughter and jokes, teamwork, es tired for lack of time to work but I am still grateful. Fighting friendships dissolve and become a family. Although sometime always Sumbawagen.<br />
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<br><br />
<br><br />
<br><br />
<br />
Gita Feny Lestari<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/f/fc/481px-Gita.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
<br />
Gita Feny lestari was born February,22th 1995, Grade 1st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
<br />
Indah Nurulita<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/9/98/Indah.png" WIDTH=100 HEIGHT=130 ><br />
<br><br />
Indah Nurulita was born March,28th 1995, Grade 1st, sumbawagen is a second home to me recently that I got a lot of experience with them, spend time with them gives a distinct impression. Although the task after task, but all come with their mild compactness feels real. spirit sumbawagen.<br />
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<br><br />
<br><br />
<br><br />
<br />
<br />
M. Al Azhar<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/2/2d/465px-Azhar.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Muhammad Al-Azhar was born june,25th 1995, Grade 1st, to be cool do not need an expert to play music, basketball or the other. enough to understand that science is cool from everything, and IGEM is a cool competition that must be followed.<br />
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<br><br />
<br><br />
<br><br />
<br />
<br />
M. Isro Al Fajri<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/a/a1/443px-Isro.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Mochammad isro alfajri was born December, 20th 1995, Grade 1st, and i especialist and leader in webdesign, humans and nature is like the driver with the vehicle. men that will bring natural conditions to welfare or destruction. because man is the leader in this world, and iGEM provides a view of nature to make it more meaningful.<br />
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<br><br />
<br><br />
<br><br />
<br />
Nugrah Analiadi Putra<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/6/6a/463px-Nugrah.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Nugrah Analiadi Putrawas born December,8th 1994, Grade 1st Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
---- <br />
<br><br />
<br><br />
<br><br />
<br />
Rian Adha Ardinata<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/8/85/428px-Rian.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Rian Adha ardinata was born May,9th 1995, Grade 1st, IGEM gives a new view to me. that life is a reciprocal relationship, whereby we must be able to utilize the natural resources to make this earth a smile.<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
Yulianti<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/b/b1/Yuli.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Yulianti was born july 1995,14th Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
Yunianti<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/f/f1/Yuni.png" WIDTH=110 HEIGHT=120 ><br />
<br><br />
<br />
Yunianti was born May,27th 1994, Grade 1st, IGEM is make my knowledge increase. IGEM makes me can to look nem other world never I look and feel.<br />
<br><br />
<br><br />
<br><br />
<br />
ここ<br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Team
Team:Sumbawagen/Team
2013-09-27T18:52:43Z
<p>Jinhee: </p>
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<h2>Advisor</h2><br />
<br><br />
Dr. Zulkieflimansyah <br />
<br><br />
<h2>instructor</h2><br />
<br><br />
Dr. Arief Budi Witarto<br />
<br><br />
Julmansyah<br />
<br><br />
<br><br />
<br><br />
<br />
<h2>Student Member</h2><br />
<br><br />
Fahmi Dwilaksono<br />
<br><br />
Abdhi Surya Perkasa<br />
<br><br />
Adelia Elviantari<br />
<br><br />
Arif Muhammad<br />
<br><br />
Asmawati<br />
<br><br />
Cendra Harri Pahlawan<br />
<br><br />
Cindy Suci Ananda<br />
<br><br />
Gita Feny Lestari<br />
<br><br />
Indah Nurulita<br />
<br><br />
M. Al Azhar<br />
<br><br />
M. Isro Al Fajri<br />
<br><br />
Nugrah Analiadi Putra<br />
<br><br />
Rian Adha Ardinata<br />
<br><br />
Yulianti<br />
<br><br />
Yunianti<br />
<br><br />
<br><br />
<br />
<h2> </h2><br />
<br />
Fahmi Dwilaksono<br />
<br><br />
<br />
<IMG SRC="https://static.igem.org/mediawiki/2013/thumb/0/0c/Fahmi.png/469px-Fahmi.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Fahmi Dwilaksono as Leader was born April, 19th 1995,Grade 1st, Joinning iGEM is a big prize for me. I know I have to work hard and study hard everyday. But I have a solid team : my lovely advisors, instructors, and of course my teammates, so I enjoy this project. I learn a lot about synthetic biology, and I realize it’s very useful for life. It would be excited to see other teams around the world, and hopefully we will get the best achievement in HongKong. Fighting!!!<br />
----<br />
<br><br />
<br><br />
<br><br />
Abdi surya perkasa<br />
<br><br />
<br />
<IMG SRC="https://static.igem.org/mediawiki/2013/4/4d/Abdi.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Abdi surya perkasa was born July, 14th 1996,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
Adelia Elviantari<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/b/bb/433px-Adel.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Adelia Elviantari was born June,26th 2013,Grade 1st,I am grateful to be part of from sumbawagen iGEM team 2013. Because I get the experience that I had never experienced before during the iGEM 2013, me and my friends taught new things that open our eyes to the sophistication of the technology.<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
Arif Muhamad<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/8/86/Arif.png" WIDTH=120 HEIGHT=120 ><br />
<br><br />
<br />
Arif Muhamad was born March, 3rd 1995,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best.<br />
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Asmawati<br />
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<IMG SRC="https://static.igem.org/mediawiki/2013/3/3a/475px-Asmawati.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Asmawati was born May,8th 1995, Grade 1 st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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Cendra Harri Pahlawan<br />
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<IMG SRC="https://static.igem.org/mediawiki/2013/thumb/2/26/Cendra.png/180px-Cendra.png" WIDTH=100 HEIGHT=130 ><br />
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Cendra Hari pahlawan was born November,10th 1994, Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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Cindy Suci Ananda<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/7/72/535px-Cindy.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Cindy Suci Ananda was born September, 16th 1996, Grade 1st, get a chance to join in and become part of the Sumbawagen is a boon for me, and I am very grateful with this grace, iGEM and sumbawagen taught the meaning of many things. Thick and thin, laughter and jokes, teamwork, es tired for lack of time to work but I am still grateful. Fighting friendships dissolve and become a family. Although sometime always Sumbawagen.<br />
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Gita Feny Lestari<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/f/fc/481px-Gita.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
<br />
Gita Feny lestari was born February,22th 1995, Grade 1st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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Indah Nurulita<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/9/98/Indah.png" WIDTH=100 HEIGHT=130 ><br />
<br><br />
Indah Nurulita was born March,28th 1995, Grade 1st, sumbawagen is a second home to me recently that I got a lot of experience with them, spend time with them gives a distinct impression. Although the task after task, but all come with their mild compactness feels real. spirit sumbawagen.<br />
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M. Al Azhar<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/2/2d/465px-Azhar.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Muhammad Al-Azhar was born june,25th 1995, Grade 1st, to be cool do not need an expert to play music, basketball or the other. enough to understand that science is cool from everything, and IGEM is a cool competition that must be followed.<br />
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M. Isro Al Fajri<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/a/a1/443px-Isro.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Mochammad isro alfajri was born December, 20th 1995, Grade 1st, and i especialist and leader in webdesign, humans and nature is like the driver with the vehicle. men that will bring natural conditions to welfare or destruction. because man is the leader in this world, and iGEM provides a view of nature to make it more meaningful.<br />
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<br />
Nugrah Analiadi Putra<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/6/6a/463px-Nugrah.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Nugrah Analiadi Putrawas born December,8th 1994, Grade 1st Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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Rian Adha Ardinata<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/8/85/428px-Rian.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Rian Adha ardinata was born May,9th 1995, Grade 1st, IGEM gives a new view to me. that life is a reciprocal relationship, whereby we must be able to utilize the natural resources to make this earth a smile.<br />
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Yulianti<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/b/b1/Yuli.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Yulianti was born july 1995,14th Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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<br><br />
<br><br />
<br />
Yunianti<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/f/f1/Yuni.png" WIDTH=110 HEIGHT=120 ><br />
<br><br />
<br />
Yunianti was born May,27th 1994, Grade 1st, IGEM is make my knowledge increase. IGEM makes me can to look nem other world never I look and feel.<br />
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ここ<br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Diary
Team:Tokyo-NoKoGen/Diary
2013-09-27T18:47:50Z
<p>Jinhee: </p>
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<br />
<div id="wrapper"><br />
<br />
<div id="title_logo"> <br />
<br />
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<a href="https://2013.igem.org"><img class="iGEM" src="https://static.igem.org/mediawiki/2013/5/58/IGEM.png"></a> <br />
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<td class="home"><a href="https://2013.igem.org/Team:Tokyo-NoKoGen"><span class="home"></span></a></td><br />
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<br />
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<br />
<div id="index"><br />
<br />
<br />
<h1 id="index_title"></h1><br />
<ul id="contents"><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Diary"><li><a href="#diary"><strong>Diary</a></strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol"><li><strong>Protocol<strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Safety"><li><strong>Safety</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement"><li><strong>Acknowledgement</strong></li><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Attribution"><li><strong>Attribution</strong></a></li><br />
<br />
</ul><br />
</div><br />
<br />
<div id="main"><br />
<br />
<BR><br />
<h1 id="april">April</h1><br />
<BR><strong>18</strong><br />
<p><br />
Our first official gathering of Tokyo-NoKoGen 2013! Okay, let’s deicde on what we will do for the project this year.<br />
<BR><br />
<BR><strong>25</strong><br />
<p>Before we start, we looked up on what projects have been in presented in the past iGEM competitions. <br />
Each one of us picked a team of our interest, and made a presentation to the team.<br />
<BR><br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/3/35/Line.jpg><br />
<br />
<br />
<h1 id="may">May</h1><br />
<BR><strong>1</strong><br />
<BR>To understand how iGEM work, we made a seminar to understand the basics, for example how to use restriction enzymes, and how to use the BioBrick parts.<br />
<BR><br />
<BR><strong>16</strong><br />
<BR>Since we are going to present at Tokyo University very very soon, we need to practice and revise our poster presentation...Are we all ready?<br />
<BR><BR><strong>19</strong><br />
<BR>School festival at Tokyo University! We made a poster presentation to many high school students and parents on our last year's project. It was very interesting to explain about genetic engineering of E. coli to those who are not familiar with them! We also got to know other team members of iGEM Japan, they were people who were fun to be with.<br />
<BR><br />
<BR><strong>25</strong><br />
<BR>Homework for the next iGEM meeting - each person must look up on a recent paper and make a presentation to the team.<br />
<br />
<br />
<BR><br />
<br />
<img src=https://static.igem.org/mediawiki/2013/3/35/Line.jpg><br />
<h1 id="june">June</h1><br />
<BR><strong>6</strong><br />
<BR>Brainstorming...production of biodiesel? Make fertilisers using E. coli?<br />
<BR><br />
<BR><strong>20</strong><br />
<BR>Another brainstorming...shall we make an extended version of lux luminescence? Anyone with good idea? Since we are running out of time, we wil make it a compulsory for each person to come up with at least 3 ideas!<br />
<BR><br />
<br />
<br />
<img src=https://static.igem.org/mediawiki/2013/3/35/Line.jpg><br />
<h1 id="july">July</h1><br />
<strong>4</strong><br />
<BR>Everyone came up with many interesting ideas over the past two weeks. Hmmm....which one shall we challenge? Not so much time left.<br />
<BR><br />
<BR><strong>11</strong><br />
<BR>Brainstorming...Ooo, HHR, RNA oscillator...?<br />
<BR><br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/3/35/Line.jpg><br />
<h1 id="august">August</h1><br />
<BR><strong>14</strong><br />
<BR>Yay, we got BBa_K1053000, our first ribozyme!<br />
<BR><br />
<BR><strong>18</strong><br />
<BR>Ohki: E-mail was sent to me(Ohki). It said as follows: Oscillator group members are Ito, Sakamoto and Takabayashi (modeling). This was the first time I had to make a simulation for RNA oscillation. Hey, who decided on that?<br />
<BR><br />
<BR><strong>19</strong><br />
<BR>Ohki: I(Ohki) decided the plan roughly. First of all, I researched what kind of software there are. I also studied how they simulate the entire system, in the protein oscillation article. <br />
<BR><br />
<BR><strong>25</strong><br />
<BR>iGEM Japan Party (this time, I(Ohki) didnt simulate at all)<br />
<BR><br />
<BR><strong>27</strong><br />
<BR>Ohki: I installed the cellillustrator. This is the software which enables us to simulate biological phenomenon. But, in this software, I was not able to simulate concisely. For example, I was not able to consider the generation of complex between HHR and taRNA. Because I roughly simulated the system, the oscillation occured. This was an obvious result because it was set to be so.<br />
<BR><br />
<BR><strong>28</strong><br />
<BR>We dicided on wiki, poster and oral presentation members<br />
<BR>Ohki: Prof. Sode said to me, "parameter in this model was not enough at all." So, I decided to recreate the simulation and decided that I was going to use other software.<br />
<BR><br />
<BR><strong>30</strong><br />
<BR>Ippei, Shunsuke: We got BBa_K1053003 and BBa_K1053004<br />
<br />
<br />
<br />
<img src=https://static.igem.org/mediawiki/2013/3/35/Line.jpg><br />
<br />
<br />
<BR><br />
<BR><br />
<h1 id="september">September</h1><br />
<BR><strong>3</strong><br />
<BR>Ippei, Shunsuke: Made TOP10 competent cells<br />
<BR><br />
<BR><strong>5</strong><br />
<BR>Eri, Masaki: Construction of RNA scaffold (containing MS2/PP7 aptamer) BBa_K1053110 <br />
<BR><br />
<BR><strong>7</strong><br />
<BR>Eri, Masaki:Sequence analysis of constructed "RNA scaffold" parts. Okay, the sequence seems fine!<br />
<BR><br />
<BR><strong>9</strong><br />
<BR>Ippei, Shunsuke: Construction of pSB3C5-PBAD-HHR-GRP-DT and evaluation. Hopefully it works!<br />
<BR><br />
<BR><strong>10</strong><br />
<BR>Get ready for BioBrick submition! Woohoo<br />
<BR>Eri, Masaki: We received our synthesized genes, FA-linker-MS2 and FB-linker-PP7! Yes! <br />
<BR><br />
<BR><strong>11</strong><br />
<BR>Eri, Masaki: The three separate fragments were ligated together, however obtained no colonies. We decided to construct using another method.<br />
<BR><br />
<BR><strong>12</strong><br />
<BR>Ohki: I installed Simbiology and studied how this software works.<br />
<BR><br />
<BR><strong>13</strong><br />
<BR>Ippei, Shunsuke: RE-evaluation of pSB3C5-PBAD-HHR-GRP-DT and evaluation. Hopefully it works this time.<br />
<BR><br />
<BR><strong>16</strong><br />
<BR>Ippei, Shunsuke: We got three HHRs composing RNA oscillator and HHR fused with RNA scaffold. But oh no...they have extra <i>Xba</I>I :(<br />
<BR><br />
<BR><strong>19</strong><br />
<BR>Ohki: I simulated our RNA oscillator and noticed this system doesn't work by substituting any parameters.<br />
<BR><br />
<BR><strong>20</strong><br />
<BR>Ippei, Shunsuke: Evaluation of pSB3C5-PBad-tr(12)-HHR-GFP-DT by taking a time course. When is the optimal time to see the difference?<br />
<BR>Eri: Time to order our T-shirt!<br />
<BR><img src=https://static.igem.org/mediawiki/2013/a/aa/%EF%BC%B4%E3%82%B7%E3%83%A3~1.PNG width=300><br />
<BR>Ohki: I formularized differential equation to confirm whether oscillation doesn't occur and solve them.<br />
<BR><br />
<BR><strong>22</strong><br />
<BR>Eri, Masaki: Construction of split GFP fused with MS2/PP7 proteins, Pconst(w)-RBS-MS2-RBS-PP7-DT<br />
<BR><br />
<BR><strong>23</strong><br />
<BR>Eri, Masaki: Evaluated GFP fluorescence in <i>E. coli</i> expressing split GFP fused with proteins and RNA scaffold. But we detected no fluorescence....why?<br />
<BR>Yukimi, Chihiro: Evalution of light sensors. Seems to behave differently under light and under dark!<br />
<BR><br />
<BR><strong>26</strong><br />
<BR>Evaluation of pSB1A3-PBAD-tr(42)-HHR-taR(12)-DT-Pconst(H)-taR(42)-DT<br />
<BR>Ohki: Time to finish writing the wiki.<br />
<BR><br />
<BR><strong>27</strong><br />
<BR>Wiki FREEZE<br />
<BR><br />
<BR><br />
<BR><img src=https://static.igem.org/mediawiki/2013/3/35/Line.jpg><br />
<h1 id="october">October</h1><br />
<BR><strong>5</strong><br />
<BR><font size=7 color=red>Asia Regional Jamboree</font><br />
<BR><br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/8/81/DSC01616.JPG width=800><br />
<BR><br />
<img src=https://static.igem.org/mediawiki/2013/5/59/DSC01611.JPG width=800><br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Protocol
Team:Tokyo-NoKoGen/Protocol
2013-09-27T18:45:46Z
<p>Jinhee: </p>
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<title>Team:Tokyo-NoKoGen - 2013.igem.org</title><br />
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<h1 id="index_title"></h1><br />
<ul id="contents"><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Diary"><li><a href="#diary"><strong>Diary</a></strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol"><li><strong>Protocol<strong></li></a><br />
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<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement"><li><strong>Acknowledgement</strong></li><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Attribution"><li><strong>Attribution</strong></a></li><br />
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<div id="main"><br />
<font size=7>Protocols</font><br />
<BR><br />
<BR><br />
<h1 id="LB"><br />
LB medium and LB agar gel<br />
Medium for cultivation of E. coli<br />
</h1><br />
<br />
<h3>LB medium (1 L)</h3><br />
1; Add about 900 mL of distilled water to beaker.<br><br />
2; Add 25 g of LB medium, Miller(MERCK) and stir. <br><br />
3; Add distilled water up to 1 L and take LB medium to media bottle.<br><br />
4; Autoclave for 20 min at 120°C.<br><br />
<br />
<h3>LB agar gel (1 L)</h3><br />
1; Prepare LB medium without autoclave (Steps 1-3 of 1L scale of LB medium).<br><br />
2; Add 15 g of agar and stirrer bar.<br><br />
3; Autoclave for 20 minutes at 120°C.<br><br />
4; Stir and cool LB medium with agar, add appropriate antibiotic (table).<br><br />
5; Pour LB medium (Step 4) in plate and cool down in clean bench.<br><br />
<br><br />
<br />
<table border="1"><br />
<td width="150"></td><br />
<td align="right" width="150">f.c.</td><tr><br />
<td width="150">Ampicillin</td><br />
<td align="right" width="150">100 μg/mL</td><tr><br />
<td width="150">Kanamycin</td><br />
<td align="right" width="150">50 μg/mL</td><tr><br />
<td width="150">Chloramphenicol </td><br />
<td align="right" width="150">30 μg/mL</td><tr><br />
<td width="150">Tetlacycline </td><br />
<td align="right" width="150">11 μg/mL</td><br />
</table><br><br><br />
<br />
<h1 id="transformation">Transformation</h1><br />
<h3>Inserting plasmid into <i>E. coli</i></h3><br />
1; Incubate frozen competent cell (DH5α) on the ice for a few minutes. <br><br />
2; Add 1~5 μL of plasmid to competent cell (DH5α) on the ice.<br><br />
3; Incubate for 20 – 30 minutes on the ice.<br><br />
4; Incubate for 45 seconds at 42°C.<br><br />
5; Add 1 mL LB medium and cultivate for 1 hour at 37°C.<br><br />
6; Spread culture medium on LB agar plate with appropriate antibiotic.<br><br><br />
<br />
<h1 id="plasmid">Plasmid extraction</h1><br />
<h3>Preparation of plasmid extracted from <i>E. coli</i></h3><br />
<br />
1; Pick up single colony from agar plate and cultivate it in 1.5 mL LB medium containing appropriate antibiotic overnight at 37°C.<br><br />
2; Move the culture medium to 1.5 mL tube.<br><br />
3; Centrifuge for 5 seconds at 15,000×g and 4 °C and discard supernatant.<br><br />
4; Add 100 μL Solution 1 to the pellet and resuspend and incubate for 3 minutes.<br><br />
5; Add 100 μL Solution 2, invert tube gently 5 times and incubate for 3 minutes.<br><br />
6; Add 100 μL Solution 3, invert tube 5 times and incubate for 3 minutes. <br><br />
8; Add 200 μL Solution 4 and invert tube 5 times and centrifuge for 3 minutes at 15,000×g and 4 °C.<br><br />
10; Take supernatant to new 1.5 mL tube and centrifuge for 3 minutes at 15,000×g and 4 °C.<br><br />
11; Vortex Bind mix for 1 min and add 800 μL Bind mix to new 1.5 mL tube.<br><br />
12; Add 400 μL supernatant after centrifugation (Step 10) to tube containing Bind mix (Step 11) and mix.<br><br />
13; Incubate for 3 minutes, centrifuge for 3 seconds at 5,000×g and 4 °C, and discard supernatant.<br><br />
14; Add 1 mL of 50% ethanol and resuspend. <br><br />
15; Centrifuge for 3 seconds at 5,000×g and 4 °C and discard supernatant.<br><br />
16; Repeat wash (Steps 14-15).<br><br />
17; Dry pellet for a few minuet under a vacuum to remove residual ethanol.<br><br />
18; Add 50 μL nuclease-free water or TE buffer and incubate for 3 minutes at 65°C.<br><br />
19; Centrifuge for 3 minutes at 15,000×g and 4 °C.<br><br />
19; 40 μL of supernatant into new 500 μL tube.<br><br><br />
<br />
<h1 id="restriction">Restriction enzyme digestion of DNA</h1><br />
<h3>Cleavage of insert DNA from plasmid</h3><br />
1; Mix DNA and restriction enzyme (Table).<br><br />
2; Incubate for 2 hours at 37°C.<br><br />
3; Incubate for 10 minutes at 65°C.<br><br />
4; Confirm the band of DNA by agar gel electrophoresis.<br><br />
<br><br />
<br><br />
<table border="1"><br />
<td width="150">reagent name</td><br />
<td align="right" width="150">volume</td><tr><br />
<td width="150"><br />
DNA<br><br />
restriction enzyme A<br><br />
restriction enzyme B<br><br />
buffre<br />
MQ</td><br />
<td align="right" width="150"><br />
5 μL<br><br />
0.5 μL<br><br />
0.5 μL<br><br />
1.5 μL<br><br />
7.5 μL<br />
</td><tr><br />
<td width="150">total</td><br />
<td align="right" width="150">15 μL</td><br />
</table><br><br><br />
<br />
<h3>Confirmation and separation of digested DNA</h3><br />
<h4>Preparation of agar gel</h4><br />
1; Add 1 g of agar to 100 mL of 1×TAE.<br><br />
2; Boil and stir until solution is dissolved and clear.<br><br />
3; Cool down, pour into container to set its shape.<br><br />
4; Wait until gel dries.<br><br />
5; Store gel in 1×TAE.<br><br><br><br />
<br />
<h1 id=electro>Agar gel electrophoresis</h1><br />
1; Place agar gel and pour 1×TAE in electrophoresis chamber.<br><br />
2; Load DNA ladder and DNA sample mixed with loading dye on agar gel.<br><br />
3; Electrophorese for 20 minutes at 100 V.<br><br />
4; Stain gel by Sybr SafeTM Gel Stain (Invitrogen).<br><br />
5; Visualize the band of DNA using UV light.<br><br />
6; Confirm the length of digested DNA.<br><br><br />
<br />
<h1 id=pcr>PCR</h1><br />
1; Add 25 μL of reagent solution (Table 1) to PCR tube.<br><br />
2; Amplify target DNA with PCR program (Table 2).<br><br />
3; Confirm the band of DNA by agar gel electrophoresis.<br><br />
<br><br />
<table border="1"><br />
<td width="350">reagent name</td><br />
<td align="right" width="100">volume</td><tr><br />
<td width="350"><br />
primeSTAR® GXL DNA polymerase<br><br />
5X GXL buffer<br><br />
dNTP<br><br />
template DNA<br><br />
fowerd primer<br><br />
reverse primer<br><br />
MQ<br />
</td><br />
<td align="right" width="100"><br />
0.5 μL<br><br />
5 μL<br><br />
2 μL<br><br />
1 μL<br><br />
1 μL<br><br />
1 μL<br><br />
14.5 μL<br />
</td><tr><br />
<td width="350">total</td><br />
<td align="right" width="100">25 μL</td><br />
</table><br />
Table 1<br />
<br><br />
<br />
<table border="1"><br />
<td width="60"></td><br />
<td width="100">Step 1</td><br />
<td width="200">Step 2</td><br />
<td width="120">Step 3</td><tr><br />
<td width="60">Cycle</td><br />
<td width="100">1</td><br />
<td width="200">30</td><br />
<td width="120">1</td><tr><br />
<td width="60"></td><br />
<td width="100" height="150" align="middle" valign="top">98℃<br><br />
1:00</td><br />
<td width="200" valign="top"><br />
&nbsp;&nbsp;&nbsp;<br />
98℃<br><br />
&nbsp;&nbsp;&nbsp;<br />
0:10<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
68℃<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
55℃<br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
4:00<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
0:10<br />
</td><br />
<td width="120" valign="top"><br><br><br />
&nbsp;&nbsp;&nbsp;68℃<br><br />
&nbsp;&nbsp;&nbsp;2:00<br><br><br><br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;4℃<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;∞</td><br />
</table><br />
Table2<br><br><br />
<br />
<br />
<br />
<h1 id=gel>Gel purification</h1><br />
<h3>Purification of DNA from agar gel</h3><br />
<h4>GENECLEAN® II Kit(NaI、glass milk、NEW Wash)/Qbiogene</h4><br />
1; Cut the objective band in the agar gel after electrophoresis and stain with SYBR Safe.<br><br />
2; Put the gel including objective DNA into 1.5 mL tube and measure the mass of that.<br><br />
3; Add 2.5-3 fold volume NaI solution into the tube (Gel: NaI =1 mg : 1 μL ).<br><br />
4; Incubate the gel at 50°C for 5 minute.<br><br />
5; Add 10 μL of glass milk and vortex.<br><br />
6; Incubate for 5 minutes and vortex per a minute.<br><br />
7; Centrifuge for 5 seconds at 15,000×g and 4°C and discard the supernatant.<br><br />
8; Add the 500μL of New Wash and resuspend.<br><br />
9; Centrifuge for 5 seconds at 15,000×g and 4°C.<br><br />
10; Repeat wash (Steps 8-9).<br><br />
11; Dry the pellet for 5-10 minutes under vacuum.<br><br />
12. Add 20 μL of nuclease-free water and resuspend.<br><br />
13. Centrifuge for 5 seconds at 15,000×g and 25°C.<br><br />
14. Transfer supernatant including objective DNA into new tube.<br><br><br />
<br />
<h1 id=ligation>Ligation</h1><br><br />
<h4>Ligation inset DNA and vector<br><br />
DNA Ligation kit Ver 2.1(SolutionⅠ)/Takara<br></h4><br />
1; Mix the insert DNA, vector and solution I (Table).<br><br />
2; Incubation at 16°C for 30 minute.<br><br />
3; Transform E. coli with ligation sample.<br><br />
<br><br />
<table border="1"><br />
<td width="150">reagent name</td><br />
<td align="right" width="150">volume</td><tr><br />
<td width="150"><br />
insert DNA<br><br />
vector<br><br />
solution I<br><br />
</td><br />
<td align="right" width="150"><br />
2 μL<br><br />
2 μL<br><br />
4 μL<br />
</td><tr><br />
<td width="150">total</td><br />
<td align="right" width="150">8 μL</td><br />
</table><br />
<br><br><br />
<br />
<h1 id=colony>Colony PCR</h1><br><br />
Confirmation of insert DNA in plasmid, directly doing PCR on <i>E. coli</i><br><br />
1; Add 10 μL of reagent solution (Table 1) to PCR tube.<br><br />
2; Pick up single colony from agar plate with tooth pick and sting replica plate (new LB agar plate).<br><br />
3; Put and stir toothpick to reagent solution (Step 1).<br><br />
4; Amplify insert DNA with PCR program (Table 2).<br><br />
5; Electrophorese PCR sample with agar gel.<br><br />
6; Check the band and length of insert DNA and decide the colony with insert DNA.<br><br />
<br><br />
<table border="1"><br />
<td width="350">reagent name</td><br />
<td align="right" width="100">volume</td><tr><br />
<td width="350"><br />
fowerd primer<br><br />
reverse primer<br><br />
Go taq® Green Master Mix(Promega)<br><br />
MQ<br />
</td><br />
<td align="right" width="100"><br />
0.5 μL<br><br />
0.5 μL<br><br />
5 μL<br><br />
4 μL<br />
</td><tr><br />
<td width="350">total</td><br />
<td align="right" width="100">10 μL</td><br />
</table><br />
Table 1<br />
<br><br />
<br />
<table border="1"><br />
<td width="60"></td><br />
<td width="100">Step 1</td><br />
<td width="200">Step 2</td><br />
<td width="120">Step 3</td><tr><br />
<td width="60">Cycle</td><br />
<td width="100">1</td><br />
<td width="200">30</td><br />
<td width="120">1</td><tr><br />
<td width="60"></td><br />
<td width="100" height="150" align="middle" valign="top">95℃<br><br />
1:00</td><br />
<td width="200" valign="top"><br />
&nbsp;&nbsp;&nbsp;<br />
95℃<br><br />
&nbsp;&nbsp;&nbsp;<br />
0:10<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
72℃<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
55℃<br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
4:00<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
0:10<br />
</td><br />
<td width="120" valign="top"><br><br><br />
&nbsp;&nbsp;&nbsp;72℃<br><br />
&nbsp;&nbsp;&nbsp;2:00<br><br><br><br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;4℃<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;∞</td><br />
</table><br />
Table2<br><br><br />
<br />
<h1 id=sequence>Sequence analysis</h1><br />
Identification of insert DNA<br> <br />
*Preparation of PCR product<br> <br />
Big Dye® Terminator Cycle Sequencing Kit Ver. 3.1 (Premix, Buffer) / Applied Biosystems<br><br />
1; Add reagent solution (Table 1) to PCR tube and amplify insert DNA with PCR program<br> (Table2).<br />
<br />
*Purification of PCR product and sequence analysis<br><br />
Agencourt CleanSEQ® and 96 R ring Super Magnetic Plate® / Beckman Coulter<br><br />
1; Add 10 μL of Agencourt CleanSEQ® 10 µL to PCR product.<br><br />
2; Add 62 μL of 85% ethanol, mix and incubate for 3 minutes.<br><br />
3; Incubate for 3 minutes on 96 R ring Super Magnetic Plate® and discard supernatant.<br><br />
4; Add 100 μL of 85% ethanol and mix.<br><br />
5; Incubate for 3 minutes on 96 R ring Super Magnetic Plate® and discard supernatant.<br><br />
6; Repeat wash (Steps 4-5).<br><br />
7; Dry for 10 minutes.<br><br />
8; Add 40 μL nuclease-free water and mix.<br><br />
9; Transfer 30μL of clear sample into a new plate for loading on the detector.<br><br />
10; Load sample on sequencer and analyze.<br><br />
<br><br />
<table border="1"><br />
<td width="350">reagent name</td><br />
<td align="right" width="100">volume</td><tr><br />
<td width="350"><br />
plasmid<br><br />
primer<br><br />
premix<br><br />
buffer<br><br />
MQ<br />
</td><br />
<td align="right" width="100"><br />
3 μL<br><br />
0.5 μL<br><br />
0.5 μL<br><br />
4 μL<br><br />
12 μL<br />
</td><tr><br />
<td width="350">total</td><br />
<td align="right" width="100">20 μL</td><br />
</table><br><br />
Table 1<br><br><br />
<br />
<table border="1"><br />
<td width="60"></td><br />
<td width="100">Step 1</td><br />
<td width="200">Step 2</td><br />
<td width="120">Step 3</td><tr><br />
<td width="60">Cycle</td><br />
<td width="100">1</td><br />
<td width="200">30</td><br />
<td width="120">1</td><tr><br />
<td width="60"></td><br />
<td width="100" height="150" align="middle" valign="top">95℃<br><br />
1:00</td><br />
<td width="200" valign="top"><br />
&nbsp;&nbsp;&nbsp;<br />
95℃<br><br />
&nbsp;&nbsp;&nbsp;<br />
0:10<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
60℃<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
50℃<br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
4:00<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br />
0:10<br />
</td><br />
<td width="120" valign="top"><br><br><br />
&nbsp;&nbsp;&nbsp;60℃<br><br />
&nbsp;&nbsp;&nbsp;2:00<br><br><br><br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;4℃<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;∞</td><br />
</table><br><br />
Table2<br />
<br />
<br><br></tr><br />
</tr><br />
<tr class="f-sp"><br />
<td><img src="Resources/_clear.gif" border=0 width=6 height=1 alt="" style="float:left"></td><br />
<td><img src="Resources/_clear.gif" border=0 width=239 height=1 alt="" style="float:left"></td><br />
<td height=1><img src="Resources/_clear.gif" border=0 width=1 height=1 alt="" style="float:left"></td><br />
</tr><br />
</table><br />
</div><br />
<br />
</div><br />
<br />
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<li class="IK"><a target="_blank" href="http://www.tuat.ac.jp/~kakusan/index.html"><img src="https://static.igem.org/mediawiki/2013/8/87/池袋研究室.png"></a></li><br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Safety
Team:Tokyo-NoKoGen/Safety
2013-09-27T18:41:44Z
<p>Jinhee: </p>
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<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Diary"><li><a href="#diary"><strong>Diary</a></strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol"><li><strong>Protocol<strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Safety"><li><strong>Safety</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement"><li><strong>Acknowledgement</strong></a></li><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Attribution"><li><strong>Attribution</strong></a></li><br />
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<p align=center><font size=7>Our Safety Form</font></p><br />
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<BR><font size=4><br />
<span style="background-color:#ffff66;">Q.Do the biological materials used in your lab work pose any of the following risks? </span></p><br />
<p><B>a. Risks to the safety and health of team members or others working in the lab?</p></b><br />
No, the biological materials used in our lab work do not pose any risks to the safety and health of lab members. We perform all experiments fully equipped with safety glasses, lab coat and gloves. We do not treat any toxic chemicals or subbstances. All experiments are performed in safety level 2 room.<br />
<BR><br />
<p><B>b. Risks to the safety and health of the general public, if released by design or by accident?</p></b><br />
No, the biological materials used in our lab work do not pose any risks to the safety and health of lab members. The only species we use is <I>Escherichia coli</I>, all of which are classified as risk group 1. They do not harm human health, and will not survive long outside the laboratory.<br />
<BR><br />
<p><B>c. Risks to the environment, if released by design or by accident?</p></b><br />
No, the biological materials used in our lab work do not pose any risks to the environment. <I>Escherichia coli</I> that we handle will not survive long out side the laboratory.<br />
<BR><br />
<p><B>d. Risks to security through malicious misuse by individuals, groups, or countriest?</p></b><br />
Our BioBrick part cannot be directly used for malicious misuse, because our aim for this year's project is to create a light oscillator, which only uses naturally existing proteins such as GFP and luciferase.<br />
<BR><br />
<BR><br />
<span style="background-color:#ffff66;">Q.If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? </span><br />
<p>We used <i>E. coli</I> (strain B and K12) as main chassis organisms and there are instruments and method to deal with large scale of these <i>E. coli</I>. So, there will be little risks if we use our <i>E. coli</i> as a commercial/industrial product.</p><br />
<BR><br />
<BR><br />
<span style="background-color:#ffff66;">Q.Does your project include any design features to address safety risks? </span></p><br />
<p>Our constructed recombinant <I>E. coli</I> must be treated in dark condition, because it has some light sensors, so we are trying to make the recombinant <i>E. coli</I> die when it is exposed to sunlight (UV light). If these <i>E. coli</I> escape from the lab (dark condition), it will die.<br />
<BR><br />
<BR><br />
<span style="background-color:#ffff66;">Q.What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution's safety training requirements, if available. </span></p><br />
<p>Our lab group holds safety meeting every Monday. In the meeting, we report any danger that arose in a week. We record it on our website but we cannot provide you the URL due to our data security.<br />
<BR><br />
<BR><br />
<span style="background-color:#ffff66;">Q.Under what biosafety privision will/do you work? </span></p><br />
<p><B>a. Please provide a link to your institution biosafety guidelines</p></b><br />
http://www.tuat.ac.jp/~idenshi/Japanese%20Files/Kitei_Folder/kitei.html<br />
<BR><br />
<p><B>b. Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you amde to your project plan based on their review</p></b><br />
We have our own Instutitional Biosafety Committee and as our past iGEM teams, we submitted report about our experiment. Our instructors are monitoring our experiments closely, and they have not identified any safety issues.<br />
<BR><br />
<p><B>c. Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible</p></b><br />
http://www.lifescience.mext.go.jp/bioethics/anzen.html#kumikae<br />
<BR><br />
<p><B>d. According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab?</p></b><br />
According to the manual, our lab has the room that meets the requirements for biosafety level 2.<br />
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<u>Safety forms were approved on September 17 2013, by Kelly Drinkwater.</u><br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement
Team:Tokyo-NoKoGen/Acknowledgement
2013-09-27T18:41:17Z
<p>Jinhee: </p>
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<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Diary"><li><a href="#diary"><strong>Diary</a></strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol"><li><strong>Protocol<strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Safety"><li><strong>Safety</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement"><li><strong>Acknowledgement</strong></a></li><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Attribution"><li><strong>Attribution</strong></a></li><br />
<br />
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<BR><br />
<BR><br />
<p align=center><font size=7>Acknowledgement</font></p><br />
<BR><br />
<BR><br />
<BR><font size=4><br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Notebook
Team:Tokyo-NoKoGen/Notebook
2013-09-27T18:40:23Z
<p>Jinhee: </p>
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<h1 id="index_title"></h1><br />
<ul id="contents"><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Diary"><li><a href="#diary"><strong>Diary</a></strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol"><li><strong>Protocol<strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Safety"><li><strong>Safety</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement"><li><strong>Acknowledgement</strong></li><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Attribution"><li><strong>Attribution</strong></a></li><br />
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<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Diary>Diary</a></p><br />
<BR><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol>Protocol</a></p><br />
<BR><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Safety>Safety</a></p><br />
<BR><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement>Acknowledgement</a></p><br />
<br><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Attribution><strong>Attribution</strong></a></p><br />
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<li class="NK"><a target="_blank" href="http://www.tuat.ac.jp/en/index.html"><img class ="NK"src="https://static.igem.org/mediawiki/2013/7/76/東京農工大学.png"></a></li><br />
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</body><br />
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</html></div>
Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Notebook
Team:Tokyo-NoKoGen/Notebook
2013-09-27T18:39:55Z
<p>Jinhee: </p>
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<h1 id="index_title"></h1><br />
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<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Diary"><li><a href="#diary"><strong>Diary</a></strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol"><li><strong>Protocol<strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Safety"><li><strong>Safety</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement"><li><strong>Acknowledgement</strong></li><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Attribution"><li><strong>Attribution</strong></a></li><br />
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<BR><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol>Protocol</a></p><br />
<BR><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Safety>Safety</a></p><br />
<BR><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement>Acknowledgement</a></p><br />
<br><br />
<p align=center><a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Attribution"><li><strong>Attribution</strong></a></p><br />
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</div><br />
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</html></div>
Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Notebook
Team:Tokyo-NoKoGen/Notebook
2013-09-27T18:39:23Z
<p>Jinhee: </p>
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<h1 id="index_title"></h1><br />
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<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Diary"><li><a href="#diary"><strong>Diary</a></strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol"><li><strong>Protocol<strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Safety"><li><strong>Safety</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement"><li><strong>Acknowledgement</strong></li><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Attribution"><li><strong>Attribution</strong></a></li><br />
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<BR><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol>Protocol</a></p><br />
<BR><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Safety>Safety</a></p><br />
<BR><br />
<p align=center><a href=https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement>Acknowledgement</a></p><br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Safety
Team:Tokyo-NoKoGen/Safety
2013-09-27T18:32:42Z
<p>Jinhee: </p>
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<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Diary"><li><a href="#diary"><strong>Diary</a></strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Protocol"><li><strong>Protocol<strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Safety"><li><strong>Safety</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement"><li><strong>Acknowledgement</strong></a></li><br />
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<p align=center><font size=7>Our Safety Form</font></p><br />
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<span style="background-color:#ffff66;">Q.Do the biological materials used in your lab work pose any of the following risks? </span></p><br />
<p><B>a. Risks to the safety and health of team members or others working in the lab?</p></b><br />
No, the biological materials used in our lab work do not pose any risks to the safety and health of lab members. We perform all experiments fully equipped with safety glasses, lab coat and gloves. We do not treat any toxic chemicals or subbstances. All experiments are performed in safety level 2 room.<br />
<BR><br />
<p><B>b. Risks to the safety and health of the general public, if released by design or by accident?</p></b><br />
No, the biological materials used in our lab work do not pose any risks to the safety and health of lab members. The only species we use is <I>Escherichia coli</I>, all of which are classified as risk group 1. They do not harm human health, and will not survive long outside the laboratory.<br />
<BR><br />
<p><B>c. Risks to the environment, if released by design or by accident?</p></b><br />
No, the biological materials used in our lab work do not pose any risks to the environment. <I>Escherichia coli</I> that we handle will not survive long out side the laboratory.<br />
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<p><B>d. Risks to security through malicious misuse by individuals, groups, or countriest?</p></b><br />
Our BioBrick part cannot be directly used for malicious misuse, because our aim for this year's project is to create a light oscillator, which only uses naturally existing proteins such as GFP and luciferase.<br />
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<BR><br />
<span style="background-color:#ffff66;">Q.If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? </span><br />
<p>We used <i>E. coli</I> (strain B and K12) as main chassis organisms and there are instruments and method to deal with large scale of these <i>E. coli</I>. So, there will be little risks if we use our <i>E. coli</i> as a commercial/industrial product.</p><br />
<BR><br />
<BR><br />
<span style="background-color:#ffff66;">Q.Does your project include any design features to address safety risks? </span></p><br />
<p>Our constructed recombinant <I>E. coli</I> must be treated in dark condition, because it has some light sensors, so we are trying to make the recombinant <i>E. coli</I> die when it is exposed to sunlight (UV light). If these <i>E. coli</I> escape from the lab (dark condition), it will die.<br />
<BR><br />
<BR><br />
<span style="background-color:#ffff66;">Q.What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution's safety training requirements, if available. </span></p><br />
<p>Our lab group holds safety meeting every Monday. In the meeting, we report any danger that arose in a week. We record it on our website but we cannot provide you the URL due to our data security.<br />
<BR><br />
<BR><br />
<span style="background-color:#ffff66;">Q.Under what biosafety privision will/do you work? </span></p><br />
<p><B>a. Please provide a link to your institution biosafety guidelines</p></b><br />
http://www.tuat.ac.jp/~idenshi/Japanese%20Files/Kitei_Folder/kitei.html<br />
<BR><br />
<p><B>b. Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you amde to your project plan based on their review</p></b><br />
We have our own Instutitional Biosafety Committee and as our past iGEM teams, we submitted report about our experiment. Our instructors are monitoring our experiments closely, and they have not identified any safety issues.<br />
<BR><br />
<p><B>c. Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible</p></b><br />
http://www.lifescience.mext.go.jp/bioethics/anzen.html#kumikae<br />
<BR><br />
<p><B>d. According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab?</p></b><br />
According to the manual, our lab has the room that meets the requirements for biosafety level 2.<br />
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<u>Safety forms were approved on September 17 2013, by Kelly Drinkwater.</u><br />
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Jinhee
http://2013.igem.org/Team:Tokyo-NoKoGen/Safety
Team:Tokyo-NoKoGen/Safety
2013-09-27T18:31:17Z
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<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Diary"><li><a href="#diary"><strong>Diary</a></strong></li></a><br />
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<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Safety"><li><strong>Safety</strong></li></a><br />
<a href="https://2013.igem.org/Team:Tokyo-NoKoGen/Acknowledgement"><li><strong>Acknowledgement</strong></a></li><br />
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<p align=center><font size=7>Our Safety Form</font></p><br />
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<span style="background-color:#ffff66;">Q.Do the biological materials used in your lab work pose any of the following risks? </span></p><br />
<p><B>a. Risks to the safety and health of team members or others working in the lab?</p></b><br />
No, the biological materials used in our lab work do not pose any risks to the safety and health of lab members. We perform all experiments fully equipped with safety glasses, lab coat and gloves. We do not treat any toxic chemicals or subbstances. All experiments are performed in safety level 2 room.<br />
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<p><B>b. Risks to the safety and health of the general public, if released by design or by accident?</p></b><br />
No, the biological materials used in our lab work do not pose any risks to the safety and health of lab members. The only species we use is <I>Escherichia coli</I>, all of which are classified as risk group 1. They do not harm human health, and will not survive long outside the laboratory.<br />
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<p><B>c. Risks to the environment, if released by design or by accident?</p></b><br />
No, the biological materials used in our lab work do not pose any risks to the environment. <I>Escherichia coli</I> that we handle will not survive long out side the laboratory.<br />
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<p><B>d. Risks to security through malicious misuse by individuals, groups, or countriest?</p></b><br />
Our BioBrick part cannot be directly used for malicious misuse, because our aim for this year's project is to create a light oscillator, which only uses naturally existing proteins such as GFP and luciferase.<br />
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<span style="background-color:#ffff66;">Q.If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? </span><br />
<p>We used <i>E. coli</I> (strain B and K12) as main chassis organisms and there are instruments and method to deal with large scale of these <i>E. coli</I>. So, there will be little risks if we use our <i>E. coli</i> as a commercial/industrial product.</p><br />
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<span style="background-color:#ffff66;">Q.Does your project include any design features to address safety risks? </span></p><br />
<p>Our constructed recombinant <I>E. coli</I> must be treated in dark condition, because it has some light sensors, so we are trying to make the recombinant <i>E. coli</I> die when it is exposed to sunlight (UV light). If these <i>E. coli</I> escape from the lab (dark condition), it will die.<br />
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<span style="background-color:#ffff66;">Q.What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution's safety training requirements, if available. </span></p><br />
<p>Our lab group holds safety meeting every Monday. In the meeting, we report any danger that arose in a week. We record it on our website but we cannot provide you the URL due to our data security.<br />
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<span style="background-color:#ffff66;">Q.Under what biosafety privision will/do you work? </span></p><br />
<p><B>a. Please provide a link to your institution biosafety guidelines</p></b><br />
http://www.tuat.ac.jp/~idenshi/Japanese%20Files/Kitei_Folder/kitei.html<br />
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<p><B>b. Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you amde to your project plan based on their review</p></b><br />
We have our own Instutitional Biosafety Committee and as our past iGEM teams, we submitted report about our experiment. Our instructors are monitoring our experiments closely, and they have not identified any safety issues.<br />
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<p><B>c. Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible</p></b><br />
http://www.lifescience.mext.go.jp/bioethics/anzen.html#kumikae<br />
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<p><B>d. According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab?</p></b><br />
According to the manual, our lab has the room that meets the requirements for biosafety level 2.<br />
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Safety forms were approved on September 17 2013, by Kelly Drinkwater.<br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Lab_Preparation_team_activities
Team:Sumbawagen/Lab Preparation team activities
2013-09-27T17:31:52Z
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Protocols
Team:Sumbawagen/Protocols
2013-09-27T17:30:23Z
<p>Jinhee: </p>
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<p style="padding-top: 0pt; " class="Free_Form">PROTOCOL<br /></p><br />
<p class="Free_Form">1.Mix Aquades and Agar<br /></p><br />
<p class="Free_Form">2.Pour mixture into Tube (Liquid Medium) and Bottle (Solid Medium)<br /></p><br />
<p class="Free_Form">3.Close the tube with cotton and bottle with the bottle cover<br /></p><br />
<p class="Free_Form">4.Wrap the tube and the bottle that we have closed using aluminium foil<br /></p><br />
<p class="Free_Form">5.Sterilise into the pressure cooker,put 1100 ml of water into the pressure cooker<br /></p><br />
<p class="Free_Form">6.Wait for 45 minutes<br /></p><br />
<p class="Free_Form">7.Take it out and let it warm<br /></p><br />
<p class="Free_Form">8.For the liquid medium put it in refrigator<br /></p><br />
<p class="Free_Form">9.For a solid medium add antibiotic<br /></p><br />
<p class="Free_Form">10.Pour into LB plates,do it in the laminar<br /></p><br />
<p class="Free_Form">11.Wait until the agar getting solid,down the position of agar plates and wair for 24 hours<br /></p><br />
<p class="Free_Form">12.Your agar is ready to use<br /></p><br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen
Team:Sumbawagen
2013-09-27T17:29:25Z
<p>Jinhee: </p>
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E. coli which able to measure the level of sugar in honey by emitting light<br />
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Glucose and fructose are major sugar component of honey. Sumbawa honey is famous as central production of forest honey in Indonesia. Sumbawa honey is also protected as geographical indication by Indonesian patent office. Taneoka et al (2009) has created a fusion of mutant glucose binding protein and firefly luciferase, which able to measure glucose level by emitting light - intended initially for blood glucose sensor application. In this project, we plan to create this construct in Biobrick format, and evaluate the ability of transgenic E. coli for the measurement of glucose in Sumbawa honey. Our final goal is to create a device which can be used for quality control of Sumbawa honey. The basic method is rapid detection for glucose and fructose by construct BioBrick which is fussion between Glucose Binding Protein (GBP) with Green Fluorescent Protein (GFP) and Red Fluorescent Protein (RFP). The BioBrick construction named "ECONEY”. <br />
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Team:Sumbawagen
2013-09-27T17:29:07Z
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E. coli which able to measure the level of sugar in honey by emitting light<br />
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Glucose and fructose are major sugar component of honey. Sumbawa honey is famous as central production of forest honey in Indonesia. Sumbawa honey is also protected as geographical indication by Indonesian patent office. Taneoka et al (2009) has created a fusion of mutant glucose binding protein and firefly luciferase, which able to measure glucose level by emitting light - intended initially for blood glucose sensor application. In this project, we plan to create this construct in Biobrick format, and evaluate the ability of transgenic E. coli for the measurement of glucose in Sumbawa honey. Our final goal is to create a device which can be used for quality control of Sumbawa honey. The basic method is rapid detection for glucose and fructose by construct BioBrick which is fussion between Glucose Binding Protein (GBP) with Green Fluorescent Protein (GFP) and Red Fluorescent Protein (RFP). The BioBrick construction named "ECONEY”. <br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Team
Team:Sumbawagen/Team
2013-09-27T17:28:30Z
<p>Jinhee: </p>
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<h2>Advisor</h2><br />
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Dr. Zulkieflimansyah <br />
<br><br />
<h2>instructor</h2><br />
<br><br />
Dr. Arief Budi Witarto<br />
<br><br />
Julmansyah<br />
<br><br />
<br><br />
<br><br />
<br />
<h2>Student Member</h2><br />
<br><br />
Fahmi Dwilaksono<br />
<br><br />
Abdhi Surya Perkasa<br />
<br><br />
Adelia Elviantari<br />
<br><br />
Arif Muhammad<br />
<br><br />
Asmawati<br />
<br><br />
Cendra Harri Pahlawan<br />
<br><br />
Cindy Suci Ananda<br />
<br><br />
Gita Feny Lestari<br />
<br><br />
Indah Nurulita<br />
<br><br />
M. Al Azhar<br />
<br><br />
M. Isro Al Fajri<br />
<br><br />
Nugrah Analiadi Putra<br />
<br><br />
Rian Adha Ardinata<br />
<br><br />
Yulianti<br />
<br><br />
Yunianti<br />
<br><br />
<br><br />
<br />
<h2> </h2><br />
<br />
Fahmi Dwilaksono<br />
<br><br />
<br />
<IMG SRC="https://static.igem.org/mediawiki/2013/thumb/0/0c/Fahmi.png/469px-Fahmi.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Fahmi Dwilaksono as Leader was born April, 19th 1995,Grade 1st, Joinning iGEM is a big prize for me. I know I have to work hard and study hard everyday. But I have a solid team : my lovely advisors, instructors, and of course my teammates, so I enjoy this project. I learn a lot about synthetic biology, and I realize it’s very useful for life. It would be excited to see other teams around the world, and hopefully we will get the best achievement in HongKong. Fighting!!!<br />
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<br><br />
<br><br />
Abdi surya perkasa<br />
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<br />
<IMG SRC="https://static.igem.org/mediawiki/2013/4/4d/Abdi.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Abdi surya perkasa was born July, 14th 1996,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best<br />
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<br><br />
<br><br />
<br><br />
<br />
Adelia Elviantari<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/b/bb/433px-Adel.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Adelia Elviantari was born June,26th 2013,Grade 1st,I am grateful to be part of from sumbawagen iGEM team 2013. Because I get the experience that I had never experienced before during the iGEM 2013, me and my friends taught new things that open our eyes to the sophistication of the technology.<br />
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<br><br />
<br><br />
<br><br />
<br />
Arif Muhamad<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/8/86/Arif.png" WIDTH=120 HEIGHT=120 ><br />
<br><br />
<br />
Arif Muhamad was born March, 3rd 1995,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best.<br />
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<br><br />
<br />
Asmawati<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/3/3a/475px-Asmawati.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Asmawati was born May,8th 1995, Grade 1 st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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<br><br />
<br><br />
<br><br />
<br />
Cendra Harri Pahlawan<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/thumb/2/26/Cendra.png/180px-Cendra.png" WIDTH=100 HEIGHT=130 ><br />
<br><br />
<br><br />
<br><br />
<br />
Cendra Hari pahlawan was born November,10th 1994, Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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<br><br />
<br><br />
<br><br />
<br />
Cindy Suci Ananda<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/7/72/535px-Cindy.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Cindy Suci Ananda was born September, 16th 1996, Grade 1st, get a chance to join in and become part of the Sumbawagen is a boon for me, and I am very grateful with this grace, iGEM and sumbawagen taught the meaning of many things. Thick and thin, laughter and jokes, teamwork, es tired for lack of time to work but I am still grateful. Fighting friendships dissolve and become a family. Although sometime always Sumbawagen.<br />
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<br><br />
<br><br />
<br><br />
<br />
Gita Feny Lestari<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/f/fc/481px-Gita.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
<br />
Gita Feny lestari was born February,22th 1995, Grade 1st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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<br><br />
<br><br />
<br><br />
<br />
<br />
Indah Nurulita<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/9/98/Indah.png" WIDTH=100 HEIGHT=130 ><br />
<br><br />
Indah Nurulita was born March,28th 1995, Grade 1st, sumbawagen is a second home to me recently that I got a lot of experience with them, spend time with them gives a distinct impression. Although the task after task, but all come with their mild compactness feels real. spirit sumbawagen.<br />
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<br><br />
<br><br />
<br><br />
<br />
<br />
M. Al Azhar<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/2/2d/465px-Azhar.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Muhammad Al-Azhar was born june,25th 1995, Grade 1st, to be cool do not need an expert to play music, basketball or the other. enough to understand that science is cool from everything, and IGEM is a cool competition that must be followed.<br />
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<br><br />
<br><br />
<br><br />
<br />
<br />
M. Isro Al Fajri<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/a/a1/443px-Isro.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Mochammad isro alfajri was born December, 20th 1995, Grade 1st, and i especialist and leader in webdesign, humans and nature is like the driver with the vehicle. men that will bring natural conditions to welfare or destruction. because man is the leader in this world, and iGEM provides a view of nature to make it more meaningful.<br />
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<br><br />
<br><br />
<br><br />
<br />
Nugrah Analiadi Putra<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/6/6a/463px-Nugrah.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
<br />
Nugrah Analiadi Putrawas born December,8th 1994, Grade 1st Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
---- <br />
<br><br />
<br><br />
<br><br />
<br />
Rian Adha Ardinata<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/8/85/428px-Rian.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Rian Adha ardinata was born May,9th 1995, Grade 1st, IGEM gives a new view to me. that life is a reciprocal relationship, whereby we must be able to utilize the natural resources to make this earth a smile.<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
Yulianti<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/b/b1/Yuli.png" WIDTH=100 HEIGHT=120 ><br />
<br><br />
Yulianti was born july 1995,14th Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
----<br />
<br><br />
<br><br />
<br><br />
<br />
Yunianti<br />
<br><br />
<IMG SRC="https://static.igem.org/mediawiki/2013/f/f1/Yuni.png" WIDTH=110 HEIGHT=120 ><br />
<br><br />
<br />
Yunianti was born May,27th 1994, Grade 1st, IGEM is make my knowledge increase. IGEM makes me can to look nem other world never I look and feel.<br />
<br><br />
<br><br />
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<br />
ここ<br />
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Jinhee
http://2013.igem.org/Team:Sumbawagen/Team
Team:Sumbawagen/Team
2013-09-27T17:27:52Z
<p>Jinhee: </p>
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ここ<br />
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SUMBAWA GEN<br />
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'''Advisors:'''<br />
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*''' Advisor ''': Dr. Zulkieflimansyah <br />
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*''' instructor 1 ''': Dr. Arief Budi Witarto<br />
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*''' instructor 2 ''': Julmansyah<br />
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'''Member:'''<br />
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*'''Student 1''': Fahmi Dwilaksono<br />
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*'''Student 2''': Abdhi Surya Perkasa<br />
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*'''Student 3''': Adelia Elviantari<br />
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*'''Student 4''': Arif Muhammad<br />
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*'''Student 5''': Asmawati<br />
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*'''Student 6''': Cendra Harri Pahlawan<br />
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*'''Student 7''': Cindy Suci Ananda<br />
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*'''Student 8''': Gita Feny Lestari<br />
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*'''Student 9''': Indah Nurulita<br />
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*'''Student 10''': M. Al Azhar<br />
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*'''Student 11''': M. Isro Al Fajri<br />
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*'''Student 12''': Nugrah Analiadi Putra<br />
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*'''Student 13''': Rian Adha Ardinata<br />
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*'''Student 14''': Yulianti<br />
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*'''Student 15''': Yunianti<br />
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*'''Fahmi Dwilaksono'''<br />
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[[File:Fahmi.png|110px|fahmi|left|thumb|link=http://https://igem.org/File:Fahmi.png]]<br />
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Fahmi Dwilaksono as Leader was born April, 19th 1995,Grade 1st, Joinning iGEM is a big prize for me. I know I have to work hard and study hard everyday. But I have a solid team : my lovely advisors, instructors, and of course my teammates, so I enjoy this project. I learn a lot about synthetic biology, and I realize it’s very useful for life. It would be excited to see other teams around the world, and hopefully we will get the best achievement in HongKong. Fighting!!!<br />
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*'''Abdi surya perkasa'''<br />
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[[File:abdi.png|110px|abdhi|left|thumb|link=http://https://igem.org/File:abdi.png|abdi]]<br />
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Abdi surya perkasa was born July, 14th 1996,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best<br />
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*'''Adelia Elviantari'''<br />
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[[File:adel.png|110px|adel|left|thumb|link=http://https://igem.org/File:adel.png]]<br />
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Adelia Elviantari was born June,26th 2013,Grade 1st,I am grateful to be part of from sumbawagen iGEM team 2013. Because I get the experience that I had never experienced before during the iGEM 2013, me and my friends taught new things that open our eyes to the sophistication of the technology.<br />
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*'''Arif Muhamad'''<br />
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[[File:arif.png|110px|arif|left|thumb|link=http://https://igem.org/File:arif.png|arif]]<br />
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Arif Muhamad was born March, 3rd 1995,Grade 1st, Biotechnology is the science that the public welfare, and IGEM is a very good competition to develop the science of biotechnology. because God has given us all the natural resources that we should be utilizing the best.<br />
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*'''Asmawati'''<br />
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[[File:asmawati.png|110px|asma|left|thumb|link=http://https://igem.org/File:asma.png|asmawati]]<br />
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Asmawati was born May,8th 1995, Grade 1 st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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*''' Cendra Harri Pahlawan'''<br />
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[[File:cendra.png|110px|cendra|left|thumb|link=http://https://igem.org/File:cendra.png|cendra]]<br />
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Cendra Hari pahlawan was born November,10th 1994, Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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*''' Cindy Suci Ananda'''<br />
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[[File:cindy.png|110px|cindy|left|thumb|link=http://https://igem.org/File:cindy.png|cindy]]<br />
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Cindy Suci Ananda was born September, 16th 1996, Grade 1st, get a chance to join in and become part of the Sumbawagen is a boon for me, and I am very grateful with this grace, iGEM and sumbawagen taught the meaning of many things. Thick and thin, laughter and jokes, teamwork, es tired for lack of time to work but I am still grateful. Fighting friendships dissolve and become a family. Although sometime always Sumbawagen.<br />
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*''' Gita Feny Lestari'''<br />
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[[File:gita.png|110px|gita|left|thumb|link=http://https://igem.org/File:gita.png|gita]]<br />
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Gita Feny lestari was born February,22th 1995, Grade 1st, man is a leader in the world, therefore man should keep this natural order to remain sustainable with memanfatkannya effectively, because our earth is getting old. IGEM gives us the opportunity to help the well-being of nature.<br />
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*''' Indah Nurulita'''<br />
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[[File:indah.png|110px|indah|left|thumb|link=http://https://igem.org/File:indah.png|Indah]]<br />
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Indah Nurulita was born March,28th 1995, Grade 1st, sumbawagen is a second home to me recently that I got a lot of experience with them, spend time with them gives a distinct impression. Although the task after task, but all come with their mild compactness feels real. spirit sumbawagen.<br />
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*''' M. Al Azhar'''<br />
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[[File:azhar.png|110px|azhar|left|thumb|link=http://https://igem.org/File:azhar.png|Azhar]]<br />
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Muhammad Al-Azhar was born june,25th 1995, Grade 1st, to be cool do not need an expert to play music, basketball or the other. enough to understand that science is cool from everything, and IGEM is a cool competition that must be followed.<br />
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*''' M. Isro Al Fajri'''<br />
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[[File:Isro.png|110px|Isro|left|thumb|link=http://https://igem.org/File:Isro.png|Isro]]<br />
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Mochammad isro alfajri was born December, 20th 1995, Grade 1st, and i especialist and leader in webdesign, humans and nature is like the driver with the vehicle. men that will bring natural conditions to welfare or destruction. because man is the leader in this world, and iGEM provides a view of nature to make it more meaningful.<br />
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*''' Nugrah Analiadi Putra'''<br />
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[[File:nugrah.png|110px|nugrah|left|thumb|link=http://https://igem.org/File:nugrah.png|nugrah]]<br />
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Nugrah Analiadi Putrawas born December,8th 1994, Grade 1st Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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*''' Rian Adha Ardinata'''<br />
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[[File:Rian.png|110px|Rian|left|thumb|link=http://https://igem.org/File:Rian.png|Rian]]<br />
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Rian Adha ardinata was born May,9th 1995, Grade 1st, IGEM gives a new view to me. that life is a reciprocal relationship, whereby we must be able to utilize the natural resources to make this earth a smile.<br />
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*''' Yulianti'''<br />
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[[File:Yuli.png|110px|Yuli|left|thumb|link=http://https://igem.org/File:Rian.png|Yuli]]<br />
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Yulianti was born july 1995,14th Grade 1st, Life is an adventure. we have to know the purpose of our life in this world, because if we do not know our purpose in this world then we will be lost, IGEM made me realize that life is full of new things that have to be realized.<br />
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*''' Yunianti'''<br />
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[[File:Yuni.png|110px|Yuni|left|thumb|link=http://https://igem.org/File:Rian.png|Yuni]]<br />
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Yunianti was born May,27th 1994, Grade 1st, IGEM is make my knowledge increase. IGEM makes me can to look nem other world never I look and feel.<br />
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