http://2013.igem.org/wiki/index.php?title=Special:Contributions/Taufik.ha&feed=atom&limit=50&target=Taufik.ha&year=&month=2013.igem.org - User contributions [en]2024-03-29T06:15:18ZFrom 2013.igem.orgMediaWiki 1.16.5http://2013.igem.org/Team:UI-Indonesia/PartsTeam:UI-Indonesia/Parts2013-10-19T02:56:12Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Our Parts</span></h1><br />
<br />
<div><br />
<img src="https://static.igem.org/mediawiki/2013/d/de/UI-Parts.JPG" border="0" align="middle" width="965"> <br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/DescriptionTeam:UI-Indonesia/Description2013-10-19T02:53:34Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Project Blue Ivy</span></h1><br />
<br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<br />
<p><h1>World’s Problem Waiting to be Solved</h1><br />
<br />
<br/><br />
Tuberculosis is still one of the worst infection-related health problem. It kills approximately 2 million people annually, making it the number one killer of single infection disease. The global epidemic is growing and becoming more dangerous. Today, tuberculosis has infected approximately a third of world’s population, even after WHO declared a global emergency on tuberculosis on 1993.<br />
<br><br />
<br>Where is the problem? Is it that hard to cure Tuberculosis? No!<br />
<br><br>WHO’s data about Tuberculosis epidemiology shows that 95% cases and 98% deaths from tuberculosis infection occur in poor countries; at the same time, 78% of world’s population dont have access to proper Tuberculosis diagnostic facilities. This clearly tells us that, the real problem in eradicating Tuberculosis is that we don’t have a reliable diagnostic tool for people in poor countries. A mathematical modelling of TB epidemiology shows that if every man and woman in this planet have an access to appropriate diagnostic facility, the prevalence of the disease will decrease 30% globally. Thus, providing a cheap, portable, reliable and easy to use TB diagnostic tool is exactly what we aim to do.<br />
<br><br />
<br>Our diagnostic tool (or biosensor) consist of an antibody fragment as the sensitive element which binds to antigen 85, a specific protein produced by Mycobacterium sp., and split Beta-galactosidase as the reporter.<br />
<br />
</p><p><h1>The Split Beta-galactosidase a.k.a The Reporter </h1><br />
Our system is based on ternary complexation mediated protein complementation principle. To put it in a simpler way, it is two proteins interacting with mediation of another protein and uses a reporter protein thatis split into two fragments, called as split reporter. Each fragment is then fused to one of the interacting protein of interest. These reporter fragment are individually inactive, and will be re-activated when the two fragments interact and re-assemble into a functioning enzyme which can convert substrate into products and gives a unique signal.<br />
<br />
<img src="https://static.igem.org/mediawiki/2013/f/f2/Desc1-r.JPG" border="0" align="middle"><br />
<br><br />
</p><p>Figure 1. Ternary complexation mediated protein complementation (Source: Stains, C.I., et al. (2010), with slight modification)<br />
<br><br />
<br><br />
We choose split Beta-Galactosidase as our reporter protein based on several considerations. First, Broome et al., (2010), have shown that they were able to split the Beta-galactosidase enzyme into two fragments that are capable to re-assemble and shows stable enzymatic activity in bacteria and in mammalian cells1. Hence, we decided to adopt their split reporter system into our biosensor. <br><br />
<br>Second, The substrate for Beta-Galactosidase, Xgal, is cheaper than the substrate for some other protein that can also functioned as split reporter, for example, luciferin (substrate for luciferase enyzme). This is important, because our goal is to make a cheap detection system to be used in suburban area.<br><br />
<br>The signal produced by Beta-Galactosidase is visible so that we dont need another device to define the infection. The signal also lasts for a long time so that we can minimize the false negative that is often occurred in short time observation. <br />
</p><p><h1>The Single Chain Fragment Variable a.k.a The Sensitive Element</h1><br />
The antibody fragment (scFv) used in our detection system is selected based on the previous experiment conducted by Ferrara et al., (2012)2. They select 111 scFvs from the previously created antibody library3 using flow cytometry. From these 111 scFvs they tested 48 of them for their specificity to Ag85, also by flow cytometry, using biotinylated Ag85; myoglobin and streptavidin-Alexa-Fluor-633 were used as negative controls. Also, they were tested to their recognition for the three different Ag85components.From the result, 14 of them were then selected for ELISA test. Three of the candidates were eliminated, leaving 11 remains. The top eight clones were further tested in an 8x8 sandwich assay, in which yeast displaying all eightselected antibodies were used as Ag85 capture reagents, and testedwith all eight scFv-Fc fusions as detection reagents. To fulfill the need of our system, we need to select two scFvs which recognize two different sites on Ag85. Based on the data on the journal, we decided to choose antibody C11 and A2 for our biosensor. <br />
<br><br />
<br />
<img src="https://static.igem.org/mediawiki/2013/0/0a/Desc2.JPG" border="0" align="middle"><br />
</p><p>Figure 2. Antibody selection method (Source:Ferrara, F., et al. (2012))<br />
<br><br><br />
<br />
<h3>References:</h3><br />
<p style="margin-left:.5in;text-indent:-.5in"><br />
1. Broome, A.M., et al. (2010). Expanding the utility of Beta-galactosidase complementation: pieceby piece. Mol Pharm. 7(1): 60–74. doi:10.1021/mp900188e.</p><br />
<p style="margin-left:.5in;text-indent:-.5in"><br />
2. Ferrara, F., et al. (2012) Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies:Application to Antigen 85, a Tuberculosis Biomarker. PLoS ONE. 7(11):e49535.doi:10.1371/journal.pone.0049535. </p><br />
<p style="margin-left:.5in;text-indent:-.5in"><br />
3. Sblattero, D., Bradbury, A. (1999). Exploiting recombination in single bacteriato make large phage antibody libraries. Nature America. </p><br />
<p style="margin-left:.5in;text-indent:-.5in"><br />
4. Stains, C.I., et al. (2010). A General Approach for Receptor and Antibody-Targeted Detection of Native Proteins utilizing Split-Luciferase Reassembly. ACS Chem Biol. 5(10): 943–952. doi:10.1021/cb100143m.<br />
</p><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/DescriptionTeam:UI-Indonesia/Description2013-10-19T02:52:26Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Project Blue Ivy</span></h1><br />
<br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<br />
<p><h1>World’s Problem Waiting to be Solved</h1><br />
<br />
<br/><br />
Tuberculosis is still one of the worst infection-related health problem. It kills approximately 2 million people annually, making it the number one killer of single infection disease. The global epidemic is growing and becoming more dangerous. Today, tuberculosis has infected approximately a third of world’s population, even after WHO declared a global emergency on tuberculosis on 1993.<br />
<br><br />
<br>Where is the problem? Is it that hard to cure Tuberculosis? No!<br />
<br><br>WHO’s data about Tuberculosis epidemiology shows that 95% cases and 98% deaths from tuberculosis infection occur in poor countries; at the same time, 78% of world’s population dont have access to proper Tuberculosis diagnostic facilities. This clearly tells us that, the real problem in eradicating Tuberculosis is that we don’t have a reliable diagnostic tool for people in poor countries. A mathematical modelling of TB epidemiology shows that if every man and woman in this planet have an access to appropriate diagnostic facility, the prevalence of the disease will decrease 30% globally. Thus, providing a cheap, portable, reliable and easy to use TB diagnostic tool is exactly what we aim to do.<br />
<br><br />
<br>Our diagnostic tool (or biosensor) consist of an antibody fragment as the sensitive element which binds to antigen 85, a specific protein produced by Mycobacterium sp., and split Beta-galactosidase as the reporter.<br />
<br />
</p><p><h1>The Split Beta-galactosidase a.k.a The Reporter </h1><br />
Our system is based on ternary complexation mediated protein complementation principle. To put it in a simpler way, it is two proteins interacting with mediation of another protein and uses a reporter protein thatis split into two fragments, called as split reporter. Each fragment is then fused to one of the interacting protein of interest. These reporter fragment are individually inactive, and will be re-activated when the two fragments interact and re-assemble into a functioning enzyme which can convert substrate into products and gives a unique signal.<br />
<br />
<img src="https://static.igem.org/mediawiki/2013/f/f2/Desc1-r.JPG" border="0" align="middle"><br />
<br><br />
</p><p>Figure 1. Ternary complexation mediated protein complementation (Source: Stains, C.I., et al. (2010), with slight modification)<br />
<br><br />
<br><br />
We choose split Beta-Galactosidase as our reporter protein based on several considerations. First, Broome et al., (2010), have shown that they were able to split the Beta-galactosidase enzyme into two fragments that are capable to re-assemble and shows stable enzymatic activity in bacteria and in mammalian cells1. Hence, we decided to adopt their split reporter system into our biosensor. <br><br />
<br>Second, The substrate for Beta-Galactosidase, Xgal, is cheaper than the substrate for some other protein that can also functioned as split reporter, for example, luciferin (substrate for luciferase enyzme). This is important, because our goal is to make a cheap detection system to be used in suburban area.<br><br />
<br>The signal produced by Beta-Galactosidase is visible so that we dont need another device to define the infection. The signal also lasts for a long time so that we can minimize the false negative that is often occurred in short time observation. <br />
</p><p><h1>The Single Chain Fragment Variable a.k.a The Sensitive Element</h1><br />
The antibody fragment (scFv) used in our detection system is selected based on the previous experiment conducted by Ferrara et al., (2012)2. They select 111 scFvs from the previously created antibody library3 using flow cytometry. From these 111 scFvs they tested 48 of them for their specificity to Ag85, also by flow cytometry, using biotinylated Ag85; myoglobin and streptavidin-Alexa-Fluor-633 were used as negative controls. Also, they were tested to their recognition for the three different Ag85components.From the result, 14 of them were then selected for ELISA test. Three of the candidates were eliminated, leaving 11 remains. The top eight clones were further tested in an 8x8 sandwich assay, in which yeast displaying all eightselected antibodies were used as Ag85 capture reagents, and testedwith all eight scFv-Fc fusions as detection reagents. To fulfill the need of our system, we need to select two scFvs which recognize two different sites on Ag85. Based on the data on the journal, we decided to choose antibody C11 and A2 for our biosensor. <br />
<br><br />
<br />
<img src="https://static.igem.org/mediawiki/2013/0/0a/Desc2.JPG" border="0" align="middle"><br />
</p><p>Figure 2. Antibody selection method (Source:Ferrara, F., et al. (2012))<br />
<br><br><br />
<br />
<h3>References:</h3><br />
<p style="margin-left:.5in;text-indent:-.5in"><br />
1. Broome, A.M., et al. (2010). Expanding the utility of Beta-galactosidase complementation: pieceby piece. Mol Pharm. 7(1): 60–74. doi:10.1021/mp900188e.</p><br />
<p style="margin-left:.5in;text-indent:-.5in"><br />
2. Ferrara, F., et al. (2012) Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies:Application to Antigen 85, a Tuberculosis Biomarker. PLoS ONE. 7(11):e49535.doi:10.1371/journal.pone.0049535. </p><br />
<p style="margin-left:.5in;text-indent:-.5in"><br />
3. Sblattero, D., Bradbury, A. (1999). Exploiting recombination in single bacteriato make large phage antibody libraries. Nature America. </p><br />
<p style="margin-left:.5in;text-indent:-.5in"><br />
4. Stains, C.I., et al. (2010). A General Approach for Receptor and Antibody-TargetedDetection of Native Proteins utilizing Split-LuciferaseReassembly. ACS Chem Biol. 5(10): 943–952. doi:10.1021/cb100143m.<br />
</p><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/AttributionsTeam:UI-Indonesia/Attributions2013-10-19T02:45:13Z<p>Taufik.ha: </p>
<hr />
<div>{{:Te{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Attributions</span></h1><br />
<br />
<div style="background-color:white; border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<p>Our project is totally an undergraduate project. All the works in our group are done by undergraduate, starting from brainstorming the idea, wet work on the bench, lab troubleshooting, constructing wiki as well as sponsors and donors hunting, It has been a long project but it is totally rewarding. We have fun and learn a lot of things. All of team members have increase the lab work skills, journal reading skills as well as negotiating skills.<br />
</p><p>We would like to thank:<br />
</p><ul><br />
<li>God, the most gracious, the most merciful <br />
</li><li>Our parents for the constant support<br />
</li><li>dr. Budiman Bela, who has kindly allowed us to do our project in IHVCB Laboratory<br />
</li><li>All IHVCB staffs who was being very nice all the time we were working on our project<br />
</li><li>Aishah from Paris-Bettencourt iGEM team 2012 for kindly sharing her experience and give us some good advices for iGEM competition</li></ul><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/HumanPracticesTeam:UI-Indonesia/HumanPractices2013-10-19T02:44:54Z<p>Taufik.ha: </p>
<hr />
<div>{{:Te{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Human Practice</span></h1><br />
<br />
<div style="background-color:white; border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<br />
<br />
<h1>UI Student Synthetic Biology Group</h1><br />
<br><br />
UI Indonesia iGEM team is currently trying to establish a student synthetic biology club in Universitas Indonesia, which is the first of its kind in Indonesia. The primary goal of the club itself is to increase the academic society's awareness to the importance of synthetic biology. There will be a routine meeting to discuss about the potential, safety, application and future of synthetic biology, especially in Indonesia. The group will also plan and work on a project on summer, to be finally sent as UI’s delegation for the next iGEM competition.<br />
<br><br><br />
The secondary goal of the club is to provide stable funding for competing in iGEM competition annually. As the first iGEM team from Universitas Indonesia, we experience difficulties in finding sponsors and donors to fund our project. Thus, UI student synthetic biology group is founded to give better direction to synthetic biology research in UI, and consequently attract sponsors and donors.<br />
<br><br><br />
The goal of the club itself is to provide stable funding for competing in iGEM competition annually as well as to increase the academic society's awareness to the importance of synthetic biology.<br />
<br><br><br />
We have talked to our professors and several key persons in Universitas Indonesia and we have gained their support. We also have done a small scale publication to our peers about iGEM competition and the response is greatly beyond expectation. They would really love to work on a real research project, but they dont know where to go or where to find research funding since support for research is very scarce in Indonesia. This is where UI Student Synthetic Biology Group will play it’s role to provide lab facility and sufficient funding for research.<br />
<br><br><br><br><br />
<br />
<br />
<h1>Questionnaire Results</h1><br />
<p><img src="https://static.igem.org/mediawiki/2013/7/7e/Ui-hp1.JPG" border="0" align="middle"></p><br />
<p><img src="https://2013.igem.org/File:Ui-hp3.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/b/b5/Ui-hp5.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/8/8a/Ui-hp11.JPG" border="0" align="middle"></p><br />
<br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/SafetyTeam:UI-Indonesia/Safety2013-10-19T02:44:30Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Safety Form</span></h1><br />
<br />
<div style="background-color:white;border:2px solid;border-radius:15px;"><br />
<br><br />
<p><img src="https://static.igem.org/mediawiki/2013/7/77/UiSafety1.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/9/9e/Safety2.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/2/23/UiSafety3.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/b/be/Safety4.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/3/30/Safety5.JPG.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/4/45/UiSafety6.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/3/35/Safety7.JPG" border="0" align="middle"></p><br />
<br><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/SafetyTeam:UI-Indonesia/Safety2013-10-19T02:43:59Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Safety Form</span></h1><br />
<br />
<div style="background-color:white;border:2px solid;border-radius:15px;"><br />
<p><img src="https://static.igem.org/mediawiki/2013/7/77/UiSafety1.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/9/9e/Safety2.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/2/23/UiSafety3.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/b/be/Safety4.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/3/30/Safety5.JPG.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/4/45/UiSafety6.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/3/35/Safety7.JPG" border="0" align="middle"></p><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/SeptemberTeam:UI-Indonesia/September2013-10-19T02:43:34Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>September</span></h1><br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<br />
<br />
<h1>September 1st</h1><br />
<li>Purification of PCR omega fragment</li><br />
<li>Making gene ruler </li><br />
<li>Running the purified product </li><br />
<li>Nanodrop the purified product : 110 ng/μL</li><br />
<li>Inoculating bacteria with pBluescript KS and pSB1C3 containg alpha fragment</li><br />
<li>Spreading BL21 and Top10 on agar plates with chloramphenicol antibiotic to check the optimum chloramphenicol concentration.</li><br />
<br />
<h1>September 3rd</h1><br />
<li>Nanodrop the large scale isolation product: <br />
<ul>pBluesript KS: 69.5 ng/μL</ul><br />
<ul>Alpha fragment : 179.5 ng/μL</ul></li><br />
<li>Restricting pBluescript KS using EcoRv</li><br />
<li>Nanodrop the restricted plasmid</li><br />
<li>Making competent cells</li><br />
<br />
<h1>September 4th</h1> <br />
<li>Testing te chloramphenicol concentration</li><br />
<li>Testing the enzymatic activity from PstI</li><br />
<li>Making agar plates with different concentration of chloramphenicol</li><br />
<li>Running the result of enzymatic activity test</li><br />
<br />
<h1>September 5th</h1><br />
<li>Re-testing the enzymatic activity using different buffers</li><br />
<li>Running</li><br />
<li>Checking the chloramphenicol test result</li><br />
<li>Transforming:<br />
<ul>Omeg fragment-pBluescript KS</ul><br />
<ul>(G4S)4 -pSB1C3 (ligated 3/9)</ul><br />
<ul>Lox511-psB1C3 (ligated 3/9)</ul><br />
<ul>Lox511-psB1C3 (ligated 25/8)</ul><br />
<ul>Lox511-psB1C3 (ligated 25/8)</ul><br />
<ul>(G4S)4 -pSB1C3 (ligated 25/8)</ul><br />
<ul>pQE80L</ul><br />
<ul>Re-ligating omega fragment-pSB1C3</ul></li><br />
<br />
<h1>September 6th</h1><br />
<li>Restricting:<br />
<ul>Omega fragment + EcoRI</ul><br />
<ul>pSB1C3</ul><br />
<ul>Omega fragmetn +XbaI</ul><br />
<ul>pBluscript KS + XbaI</ul><br />
<ul>Alpha fragment + XbaI</ul></li><br />
<li>Running</li><br />
<li>Nanodrop the purified digested plasmid<br />
<ul>Omega fragment + EcoRI = 7.2 ng/μL</ul><br />
<ul>pSB1C3 = 3.8 ng/μL</ul><br />
<ul>Omega fragment + XbaI = 7.7 ng/μL</ul><br />
<ul>pBluescript KS + XbaI = 9.8 ng/μL</ul><br />
<ul>Alpha fragment + Xba I = 0.35 ng/μL</ul></li><br />
<li>Running</li><br />
<li>Inoculating bacteria with pSB1C3-alpha fragment in 100mL broth</li><br />
<br />
<h1>September 9th</h1><br />
<li>Large scale pSB1C3-alpha fragment isolation</li><br />
<li>Restricting pBluescript KS using EcoRV</li><br />
<li>Digesting linearized pSB1C3 and linearized omega fragment using PstI</li><br />
<li>Making 750mL LB Broth</li><br />
<li>Making agar plates</li><br />
<li>Digesting pSB1C3-alpha fragment</li><br />
<li>Purifying the digested using LMA</li><br />
<li>Blunting Lox511 and (G4S)4 linker</li><br />
<li>Running</li><br />
<br />
<h1>September 11th</h1><br />
<li>Small scale isolation</li><br />
<li>Running the small scale isolation product</li><br />
<br />
<h1>September 13th</h1><br />
<li>Re-run the small scale iolation product</li><br />
<li>Inoculate the bacteria containing pSB1C3=omega fragment</li><br />
<br />
<h1>September 14th</h1><br />
<li>Running small scale isolation product</li><br />
<li>PCR Colony</li><br />
<li>Large scale isolation</li><br />
<li>Re-ligating omega fragment with pSB1C3</li><br />
<br />
<h1>September 15th</h1><br />
<li>Running<br />
<ul>Digested pQE+PstI</ul><br />
<ul>PQE</ul><br />
<ul>Digested alpha fragment+PstI</ul><br />
<ul>Alpha fragment</ul><br />
<ul>Marker</ul><br />
<ul>5</ul><br />
<ul>8</ul><br />
<ul>9</ul><br />
<ul>15</ul><br />
<ul>Large scale isolation pBluescript KS-omega</ul></li><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/AugustTeam:UI-Indonesia/August2013-10-19T02:42:36Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>August</span></h1><br />
<div style="background-color:white ; border:2px solid; border-radius:15px; padding:15px 15px;"><br />
<br />
<h1>August 5th</h1><br />
<li>Performing PCR gradient for alpha fragment and omega fragment with and without linker<br />
<ul> Temperatures used for the PCR are 510C, 54.50C, 60.20C, 63.90C, 670C and 69.50C</ul></li><br />
<li>Running electrophoresis to check the result<br />
<ul> The bands were very thin, so we decided to re-PCR the alpha and omega fragment</ul></li><br />
<br />
<h1>August 6th</h1><br />
<li>Linearizing LacZ full from 3rd coloni in the replica (third transformation)<br />
<li>PCR alpha and omega fragment with and without linker<br />
<ul>Temperatures used for the PCR are 500C, 53.20C, 55.50C, 61.00C, 66.80C, and 68.40C</ul><li><br />
<li>Running electrophoresis<br />
<ul>The thickest band was shown at 53.20C for alpha fragment and at 500C for omega fragment. Hence the temperatures said above will be used for PCR using Taq HiFi</ul></li><br />
<br />
<h1>August 12th</h1><br />
<li>Running<br />
<ul>We got bands with correct length, however we got thin bands on well containing omega fragment of beta galactosidase. We decided to do large scale PCR using PFx the next day.</ul></li><br />
<br />
<h1>August 13th</h1><br />
<li>Large scale PCR (50μL) using PFx enzyme with annealing temperature at 53.20C for alpha fragment, 500C for omega fragment and omega fragment+linker<br />
<ul>Running electrophoresis <br />
<ul>We got the correct length for all fragments</ul></ul></li><br />
<br />
<h1>August 14th</h1><br />
<li>Large scale PCR (50μL) using PFx enzyme with annealing temperature at 53.20C for alpha fragment, 500C for omega fragment and omega fragment+linker<br />
<ul>Running electrophoresis:<br />
<ul>Correct length for all fragments</ul></ul><br />
<ul>Restricting pQE80L using PstI<br />
<ul>We use three tubes, each of them containing PstI with different volume: 0.5 μL, 5 μL, and without any enzyme added for negative control.</ul></ul><br />
§ This step was done in order to check the enzyme, because we are using PstI that had past its expiration date.<br />
<ul>Running the restricted PCR<br />
<ul>The result shows that the PstI is no longer in a good condition. For 1 hr incubation period, the restriction using 0.5μL PstI does not produce the correct length of pQE 80L (4751 bp)</ul></ul><br />
<ul>Incubating the restricted pQE80L overnight<br />
<ul>This step was done in order to check the activity of the enzyme in extended period.</ul></ul></li><br />
<br />
<h1>August 15th</h1> <br />
<li>Moving the pQE 80L + PstI into -400C freezer</li><br />
<li>Purifying the fragments (alpha+linker, omega, omega+llinker)from the PCR products using Low Melting Point Agarose (LMA) method.</li><br />
<br />
<h1>August 16th</h1><br />
<li>Running the incubated pQE 80L + PstI</li><br />
<br />
The enzyme works with extended time of incubation. However, star activity is also observed. Hence, we look up on the internet and found that PstI can still works for extender period of incubation up to 4 hours.<br />
<br />
<h1>August 17th</h1><br />
<li>Checking the isolated fragments' concentration using Nanodrop 2000<br />
<ul>Alpha fragment concentration: 17.1 ng/μL</ul><br />
<ul>Omega fragment concentration: 18.8 ng/μL</ul><br />
<ul>Omega+linker fragment concentration: 17.1 ng/μL</ul></li><br />
<li>Ligating pSB1C3 and omega fragment</li><br />
<br />
<h1>August 18th</h1><br />
<li>Running purified plasmid in agarose</li><br />
<li>Linearizing alpha fragment using PstI</li><br />
<li>Ligating pcDNA 3.1 + omega fragment</li><br />
<li>Ligating pcDNA 3.1 + alpha fragment</li><br />
<li>Cutting the ligated psB1C3 + omega fragment using EcoRI to verify he length of fragment inserted</li><br />
<li>Running</li><br />
<br />
<h1>August 19th</h1><br />
<li>Re-ligating psB1C3+ω</li><br />
<li>Cut pCDNA+α and pCDNA+ω with BamHI</li><br />
<li>Running</li><br />
<br />
<h1>August 20th</h1><br />
<li>Running the ligated parts</li><br />
<li>Making new competent cells</li><br />
<li>Transforming the ligated parts</li><br />
<li>Spread the transformed cells into agar plates</li><br />
<br />
<h1>August 22nd</h1><br />
<li>Making agar plates</li><br />
<li>Re-PCR alpha and omega fragments using PFX HiFi</li><br />
<li>Running electrophoresis</li><br />
<br />
<h1>August 23rd</h1><br />
<li>Ligating gBlocks to pBluescript and pSB1C3</li><br />
<li>Ligating alpha and omega fragment to pBluescript</li><br />
<li>Purify PCR products using LMA</li><br />
<li>Ligating alpha and omega fragments from PCR products to pcDNA 3.1</li><br />
<br />
<h1>August 24th</h1><br />
<li>Nanodrop the LMA product</li><br />
<li>Nanodrop the previously isolated alpha fragment</li><br />
<li>Restricting LMA with omega fragments using EcoRI</li><br />
<li>Restricting pCDNA 3.1 using EcoRI and XhoI</li><br />
<li>Restricting the previously purified alpha fragment using EcoRI</li><br />
<li>Running</li><br />
<br />
<h1>August 25th</h1><br />
<li>Restricting:<br />
<ul>LMA omega fragmetns</ul><br />
<ul>Lox511 gBlock</ul><br />
<ul>(G4S)4 gBlock</ul></li><br />
<li>Purifying the restricted plasmid from the preious day</li><br />
<li>Ligating<br />
<ul>Omega fragment-pCDNA 3.1</ul><br />
<ul>Alpha fragment-pCDNA 3.1</ul><br />
<ul>Omega fragment-pSB1C3</ul><br />
<ul>Lox511-pSB1C3</ul><br />
<ul>(G4S)4- pSB1C3</ul></li><br />
<li>Purifying the following:<br />
<ul>Omega fragment restricted with EcoRI and PstI</ul><br />
<ul>Lox511</ul><br />
<ul>(G4S)4</ul></li><br />
<br />
<h1>August 26th</h1><br />
<li>Transforming<br />
<ul>Lox511</ul><br />
<ul>(G4S)4</ul><br />
<ul>pQE 80-L</ul><br />
<ul>pQE 82-L</ul><br />
<ul>Positive control for ampicillin and chloramphenicol antibiotics</ul></li><br />
<li>Spread</li><br />
<li>Streak E.coli top10</li><br />
<li>Ligation using T4 DNA Ligase Promega + T4 NEBuffer</li><br />
<br />
<h1>August 27th</h1><br />
<li>Making replica on LB agar plate</li><br />
<li>Running the previously ligated plasmids</li><br />
<li>Restriction using BamHI and PstI</li><br />
<li>Purification of DNA</li><br />
<li>Making new agar plates</li><br />
<li>Making overnight top10</li><br />
<li>Inoculating bacteria in LB Broth for mall scale isolation</li><br />
<br />
<h1>August 28th</h1><br />
<li>PCR omega fragments</li><br />
<li>Small scale isolation</li><br />
<li>Restriction using EcoRI to check the length</li><br />
<li>Running</li><br />
<h1>August 30th</h1><br />
<li>Inoculating bacteria with pBluescript KS in 4 mL LB broth</li><br />
<li>PCR omega fragment</li><br />
<li>Making LB Agar</li><br />
<li>Making LB Broth</li><br />
<li>Making pBluescript KS replica</li><br />
<br />
<h1>August 31st</h1><br />
<li>Small scale isolaiton</li><br />
<br />
<h1>Running PCR products August 30th</h1><br />
<li>Inoculating bacteria with pBluescript KS in 4 mL LB broth</li><br />
<li>PCR omega fragment</li><br />
<li>Making LB Agar</li><br />
<li>Making LB Broth</li><br />
<li>Making pBluescript KS replica</li><br />
<br />
<h1>August 31st</h1><br />
<li>Small scale isolaiton</li><br />
<li>Running PCR products</li><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/AugustTeam:UI-Indonesia/August2013-10-19T02:42:20Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>August</span></h1><br />
<div style="background-color:whit ; border:2px solid; border-radius:15px; padding:15px 15px;"><br />
<br />
<h1>August 5th</h1><br />
<li>Performing PCR gradient for alpha fragment and omega fragment with and without linker<br />
<ul> Temperatures used for the PCR are 510C, 54.50C, 60.20C, 63.90C, 670C and 69.50C</ul></li><br />
<li>Running electrophoresis to check the result<br />
<ul> The bands were very thin, so we decided to re-PCR the alpha and omega fragment</ul></li><br />
<br />
<h1>August 6th</h1><br />
<li>Linearizing LacZ full from 3rd coloni in the replica (third transformation)<br />
<li>PCR alpha and omega fragment with and without linker<br />
<ul>Temperatures used for the PCR are 500C, 53.20C, 55.50C, 61.00C, 66.80C, and 68.40C</ul><li><br />
<li>Running electrophoresis<br />
<ul>The thickest band was shown at 53.20C for alpha fragment and at 500C for omega fragment. Hence the temperatures said above will be used for PCR using Taq HiFi</ul></li><br />
<br />
<h1>August 12th</h1><br />
<li>Running<br />
<ul>We got bands with correct length, however we got thin bands on well containing omega fragment of beta galactosidase. We decided to do large scale PCR using PFx the next day.</ul></li><br />
<br />
<h1>August 13th</h1><br />
<li>Large scale PCR (50μL) using PFx enzyme with annealing temperature at 53.20C for alpha fragment, 500C for omega fragment and omega fragment+linker<br />
<ul>Running electrophoresis <br />
<ul>We got the correct length for all fragments</ul></ul></li><br />
<br />
<h1>August 14th</h1><br />
<li>Large scale PCR (50μL) using PFx enzyme with annealing temperature at 53.20C for alpha fragment, 500C for omega fragment and omega fragment+linker<br />
<ul>Running electrophoresis:<br />
<ul>Correct length for all fragments</ul></ul><br />
<ul>Restricting pQE80L using PstI<br />
<ul>We use three tubes, each of them containing PstI with different volume: 0.5 μL, 5 μL, and without any enzyme added for negative control.</ul></ul><br />
§ This step was done in order to check the enzyme, because we are using PstI that had past its expiration date.<br />
<ul>Running the restricted PCR<br />
<ul>The result shows that the PstI is no longer in a good condition. For 1 hr incubation period, the restriction using 0.5μL PstI does not produce the correct length of pQE 80L (4751 bp)</ul></ul><br />
<ul>Incubating the restricted pQE80L overnight<br />
<ul>This step was done in order to check the activity of the enzyme in extended period.</ul></ul></li><br />
<br />
<h1>August 15th</h1> <br />
<li>Moving the pQE 80L + PstI into -400C freezer</li><br />
<li>Purifying the fragments (alpha+linker, omega, omega+llinker)from the PCR products using Low Melting Point Agarose (LMA) method.</li><br />
<br />
<h1>August 16th</h1><br />
<li>Running the incubated pQE 80L + PstI</li><br />
<br />
The enzyme works with extended time of incubation. However, star activity is also observed. Hence, we look up on the internet and found that PstI can still works for extender period of incubation up to 4 hours.<br />
<br />
<h1>August 17th</h1><br />
<li>Checking the isolated fragments' concentration using Nanodrop 2000<br />
<ul>Alpha fragment concentration: 17.1 ng/μL</ul><br />
<ul>Omega fragment concentration: 18.8 ng/μL</ul><br />
<ul>Omega+linker fragment concentration: 17.1 ng/μL</ul></li><br />
<li>Ligating pSB1C3 and omega fragment</li><br />
<br />
<h1>August 18th</h1><br />
<li>Running purified plasmid in agarose</li><br />
<li>Linearizing alpha fragment using PstI</li><br />
<li>Ligating pcDNA 3.1 + omega fragment</li><br />
<li>Ligating pcDNA 3.1 + alpha fragment</li><br />
<li>Cutting the ligated psB1C3 + omega fragment using EcoRI to verify he length of fragment inserted</li><br />
<li>Running</li><br />
<br />
<h1>August 19th</h1><br />
<li>Re-ligating psB1C3+ω</li><br />
<li>Cut pCDNA+α and pCDNA+ω with BamHI</li><br />
<li>Running</li><br />
<br />
<h1>August 20th</h1><br />
<li>Running the ligated parts</li><br />
<li>Making new competent cells</li><br />
<li>Transforming the ligated parts</li><br />
<li>Spread the transformed cells into agar plates</li><br />
<br />
<h1>August 22nd</h1><br />
<li>Making agar plates</li><br />
<li>Re-PCR alpha and omega fragments using PFX HiFi</li><br />
<li>Running electrophoresis</li><br />
<br />
<h1>August 23rd</h1><br />
<li>Ligating gBlocks to pBluescript and pSB1C3</li><br />
<li>Ligating alpha and omega fragment to pBluescript</li><br />
<li>Purify PCR products using LMA</li><br />
<li>Ligating alpha and omega fragments from PCR products to pcDNA 3.1</li><br />
<br />
<h1>August 24th</h1><br />
<li>Nanodrop the LMA product</li><br />
<li>Nanodrop the previously isolated alpha fragment</li><br />
<li>Restricting LMA with omega fragments using EcoRI</li><br />
<li>Restricting pCDNA 3.1 using EcoRI and XhoI</li><br />
<li>Restricting the previously purified alpha fragment using EcoRI</li><br />
<li>Running</li><br />
<br />
<h1>August 25th</h1><br />
<li>Restricting:<br />
<ul>LMA omega fragmetns</ul><br />
<ul>Lox511 gBlock</ul><br />
<ul>(G4S)4 gBlock</ul></li><br />
<li>Purifying the restricted plasmid from the preious day</li><br />
<li>Ligating<br />
<ul>Omega fragment-pCDNA 3.1</ul><br />
<ul>Alpha fragment-pCDNA 3.1</ul><br />
<ul>Omega fragment-pSB1C3</ul><br />
<ul>Lox511-pSB1C3</ul><br />
<ul>(G4S)4- pSB1C3</ul></li><br />
<li>Purifying the following:<br />
<ul>Omega fragment restricted with EcoRI and PstI</ul><br />
<ul>Lox511</ul><br />
<ul>(G4S)4</ul></li><br />
<br />
<h1>August 26th</h1><br />
<li>Transforming<br />
<ul>Lox511</ul><br />
<ul>(G4S)4</ul><br />
<ul>pQE 80-L</ul><br />
<ul>pQE 82-L</ul><br />
<ul>Positive control for ampicillin and chloramphenicol antibiotics</ul></li><br />
<li>Spread</li><br />
<li>Streak E.coli top10</li><br />
<li>Ligation using T4 DNA Ligase Promega + T4 NEBuffer</li><br />
<br />
<h1>August 27th</h1><br />
<li>Making replica on LB agar plate</li><br />
<li>Running the previously ligated plasmids</li><br />
<li>Restriction using BamHI and PstI</li><br />
<li>Purification of DNA</li><br />
<li>Making new agar plates</li><br />
<li>Making overnight top10</li><br />
<li>Inoculating bacteria in LB Broth for mall scale isolation</li><br />
<br />
<h1>August 28th</h1><br />
<li>PCR omega fragments</li><br />
<li>Small scale isolation</li><br />
<li>Restriction using EcoRI to check the length</li><br />
<li>Running</li><br />
<h1>August 30th</h1><br />
<li>Inoculating bacteria with pBluescript KS in 4 mL LB broth</li><br />
<li>PCR omega fragment</li><br />
<li>Making LB Agar</li><br />
<li>Making LB Broth</li><br />
<li>Making pBluescript KS replica</li><br />
<br />
<h1>August 31st</h1><br />
<li>Small scale isolaiton</li><br />
<br />
<h1>Running PCR products August 30th</h1><br />
<li>Inoculating bacteria with pBluescript KS in 4 mL LB broth</li><br />
<li>PCR omega fragment</li><br />
<li>Making LB Agar</li><br />
<li>Making LB Broth</li><br />
<li>Making pBluescript KS replica</li><br />
<br />
<h1>August 31st</h1><br />
<li>Small scale isolaiton</li><br />
<li>Running PCR products</li><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/AugustTeam:UI-Indonesia/August2013-10-19T02:41:32Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>August</span></h1><br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<br />
<h1>August 5th</h1><br />
<li>Performing PCR gradient for alpha fragment and omega fragment with and without linker<br />
<ul> Temperatures used for the PCR are 510C, 54.50C, 60.20C, 63.90C, 670C and 69.50C</ul></li><br />
<li>Running electrophoresis to check the result<br />
<ul> The bands were very thin, so we decided to re-PCR the alpha and omega fragment</ul></li><br />
<br />
<h1>August 6th</h1><br />
<li>Linearizing LacZ full from 3rd coloni in the replica (third transformation)<br />
<li>PCR alpha and omega fragment with and without linker<br />
<ul>Temperatures used for the PCR are 500C, 53.20C, 55.50C, 61.00C, 66.80C, and 68.40C</ul><li><br />
<li>Running electrophoresis<br />
<ul>The thickest band was shown at 53.20C for alpha fragment and at 500C for omega fragment. Hence the temperatures said above will be used for PCR using Taq HiFi</ul></li><br />
<br />
<h1>August 12th</h1><br />
<li>Running<br />
<ul>We got bands with correct length, however we got thin bands on well containing omega fragment of beta galactosidase. We decided to do large scale PCR using PFx the next day.</ul></li><br />
<br />
<h1>August 13th</h1><br />
<li>Large scale PCR (50μL) using PFx enzyme with annealing temperature at 53.20C for alpha fragment, 500C for omega fragment and omega fragment+linker<br />
<ul>Running electrophoresis <br />
<ul>We got the correct length for all fragments</ul></ul></li><br />
<br />
<h1>August 14th</h1><br />
<li>Large scale PCR (50μL) using PFx enzyme with annealing temperature at 53.20C for alpha fragment, 500C for omega fragment and omega fragment+linker<br />
<ul>Running electrophoresis:<br />
<ul>Correct length for all fragments</ul></ul><br />
<ul>Restricting pQE80L using PstI<br />
<ul>We use three tubes, each of them containing PstI with different volume: 0.5 μL, 5 μL, and without any enzyme added for negative control.</ul></ul><br />
§ This step was done in order to check the enzyme, because we are using PstI that had past its expiration date.<br />
<ul>Running the restricted PCR<br />
<ul>The result shows that the PstI is no longer in a good condition. For 1 hr incubation period, the restriction using 0.5μL PstI does not produce the correct length of pQE 80L (4751 bp)</ul></ul><br />
<ul>Incubating the restricted pQE80L overnight<br />
<ul>This step was done in order to check the activity of the enzyme in extended period.</ul></ul></li><br />
<br />
<h1>August 15th</h1> <br />
<li>Moving the pQE 80L + PstI into -400C freezer</li><br />
<li>Purifying the fragments (alpha+linker, omega, omega+llinker)from the PCR products using Low Melting Point Agarose (LMA) method.</li><br />
<br />
<h1>August 16th</h1><br />
<li>Running the incubated pQE 80L + PstI</li><br />
<br />
The enzyme works with extended time of incubation. However, star activity is also observed. Hence, we look up on the internet and found that PstI can still works for extender period of incubation up to 4 hours.<br />
<br />
<h1>August 17th</h1><br />
<li>Checking the isolated fragments' concentration using Nanodrop 2000<br />
<ul>Alpha fragment concentration: 17.1 ng/μL</ul><br />
<ul>Omega fragment concentration: 18.8 ng/μL</ul><br />
<ul>Omega+linker fragment concentration: 17.1 ng/μL</ul></li><br />
<li>Ligating pSB1C3 and omega fragment</li><br />
<br />
<h1>August 18th</h1><br />
<li>Running purified plasmid in agarose</li><br />
<li>Linearizing alpha fragment using PstI</li><br />
<li>Ligating pcDNA 3.1 + omega fragment</li><br />
<li>Ligating pcDNA 3.1 + alpha fragment</li><br />
<li>Cutting the ligated psB1C3 + omega fragment using EcoRI to verify he length of fragment inserted</li><br />
<li>Running</li><br />
<br />
<h1>August 19th</h1><br />
<li>Re-ligating psB1C3+ω</li><br />
<li>Cut pCDNA+α and pCDNA+ω with BamHI</li><br />
<li>Running</li><br />
<br />
<h1>August 20th</h1><br />
<li>Running the ligated parts</li><br />
<li>Making new competent cells</li><br />
<li>Transforming the ligated parts</li><br />
<li>Spread the transformed cells into agar plates</li><br />
<br />
<h1>August 22nd</h1><br />
<li>Making agar plates</li><br />
<li>Re-PCR alpha and omega fragments using PFX HiFi</li><br />
<li>Running electrophoresis</li><br />
<br />
<h1>August 23rd</h1><br />
<li>Ligating gBlocks to pBluescript and pSB1C3</li><br />
<li>Ligating alpha and omega fragment to pBluescript</li><br />
<li>Purify PCR products using LMA</li><br />
<li>Ligating alpha and omega fragments from PCR products to pcDNA 3.1</li><br />
<br />
<h1>August 24th</h1><br />
<li>Nanodrop the LMA product</li><br />
<li>Nanodrop the previously isolated alpha fragment</li><br />
<li>Restricting LMA with omega fragments using EcoRI</li><br />
<li>Restricting pCDNA 3.1 using EcoRI and XhoI</li><br />
<li>Restricting the previously purified alpha fragment using EcoRI</li><br />
<li>Running</li><br />
<br />
<h1>August 25th</h1><br />
<li>Restricting:<br />
<ul>LMA omega fragmetns</ul><br />
<ul>Lox511 gBlock</ul><br />
<ul>(G4S)4 gBlock</ul></li><br />
<li>Purifying the restricted plasmid from the preious day</li><br />
<li>Ligating<br />
<ul>Omega fragment-pCDNA 3.1</ul><br />
<ul>Alpha fragment-pCDNA 3.1</ul><br />
<ul>Omega fragment-pSB1C3</ul><br />
<ul>Lox511-pSB1C3</ul><br />
<ul>(G4S)4- pSB1C3</ul></li><br />
<li>Purifying the following:<br />
<ul>Omega fragment restricted with EcoRI and PstI</ul><br />
<ul>Lox511</ul><br />
<ul>(G4S)4</ul></li><br />
<br />
<h1>August 26th</h1><br />
<li>Transforming<br />
<ul>Lox511</ul><br />
<ul>(G4S)4</ul><br />
<ul>pQE 80-L</ul><br />
<ul>pQE 82-L</ul><br />
<ul>Positive control for ampicillin and chloramphenicol antibiotics</ul></li><br />
<li>Spread</li><br />
<li>Streak E.coli top10</li><br />
<li>Ligation using T4 DNA Ligase Promega + T4 NEBuffer</li><br />
<br />
<h1>August 27th</h1><br />
<li>Making replica on LB agar plate</li><br />
<li>Running the previously ligated plasmids</li><br />
<li>Restriction using BamHI and PstI</li><br />
<li>Purification of DNA</li><br />
<li>Making new agar plates</li><br />
<li>Making overnight top10</li><br />
<li>Inoculating bacteria in LB Broth for mall scale isolation</li><br />
<br />
<h1>August 28th</h1><br />
<li>PCR omega fragments</li><br />
<li>Small scale isolation</li><br />
<li>Restriction using EcoRI to check the length</li><br />
<li>Running</li><br />
<h1>August 30th</h1><br />
<li>Inoculating bacteria with pBluescript KS in 4 mL LB broth</li><br />
<li>PCR omega fragment</li><br />
<li>Making LB Agar</li><br />
<li>Making LB Broth</li><br />
<li>Making pBluescript KS replica</li><br />
<br />
<h1>August 31st</h1><br />
<li>Small scale isolaiton</li><br />
<br />
<h1>Running PCR products August 30th</h1><br />
<li>Inoculating bacteria with pBluescript KS in 4 mL LB broth</li><br />
<li>PCR omega fragment</li><br />
<li>Making LB Agar</li><br />
<li>Making LB Broth</li><br />
<li>Making pBluescript KS replica</li><br />
<br />
<h1>August 31st</h1><br />
<li>Small scale isolaiton</li><br />
<li>Running PCR products</li><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/JulyTeam:UI-Indonesia/July2013-10-19T02:41:09Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>July</span></h1><br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<br />
<h1>July 1st </h1><br />
<h5><br />
<p>What to do:<br />
</p><ul><br />
<li>Resuspend plasmids from the iGEM kit<br />
</li><li>Transform to E.coli cell<br />
</li><li>Store the remaining suspended plasmid<br />
<br />
</li></ul><br />
<p>Reagents and equipments<br />
</p><ul><br />
<li>Reagents<br />
<ol><br />
<li>Distilled water<br />
</li><li>Ice<br />
</li><li>50 µL competent cells for parts of choice<br />
</li><li>50 µL competent cells for positive control<br />
</li><li>Positive control<br />
</li><li>SOC Media<br />
</li><li>Agar plates with appropriate antibiotic<br />
<p> &sect; NOTE: this needs to be prepared beforehand. <br />
</p></ol><br />
</li><li>Equipments<br />
<ol><br />
<li>10 and 200 µL pipettes<br />
</li><li>Timer<br />
</li><li>Water bath (420C)<br />
<br />
<dl><br />
<dt>Procedures</dt><br />
<dd> </dd></dl><br />
</li></ol><br />
</li><li>Resuspending the part<br />
<ol><br />
<li>Mark the location of the parts using a pen or marker<br />
</li><li>Punch a hole through the foil using a pipette tip<br />
</li><li>Add 10µL of dH2O<br />
</li><li>Mix thoroughly by aspirating up and down a few times<br />
</li></ol><br />
</li><li>Inoculating the DNA<br />
<ol><br />
<li>Add 1µL of the suspended DNA to the competent cells<br />
<p> &sect; The competent cells must be kept on ice before and after the addition of the suspended DNA<br />
&sect; Make sure to mark the lid of the eppendorf tube, which tube is filled with C.C +DNA and which tube is filled with positive control.<br />
<li>Add positive control to the competent cells<br />
<p> &sect; ASK: What strain of E.coli are we going to use as the competent cell? E.coli codon plus? Top10? BL21? pLysS?<br />
<br />
</p></ol><br />
</li><li>Incubate on ice for 30 minutes<br />
<br />
</li><li>Heatshock in water bath for 1 minute<br />
<br />
</li><li>Incubate on ice for 5 min<br />
<br />
</li><li>Add 200 µL of SOC media to each transformation<br />
<p> &sect; ASK: Is it available in IHVCB? If SOC media is not available, can we replace it with regular LB broth? <br />
<br />
<li>Incubate for 2 hours at 370C<br />
<br />
</li><li>Plate out the tranformation<br />
<ol><br />
<li>Plate 20 µL to one agar plate, and the remainder to the separate plate.<br />
</li><li>Use beads to spread evenly<br />
</li><li>Incubate overnight (16hrs) at 370C<br />
<p> &sect; ASK: What antibiotics do we need? Is it available in IHVCB? <br />
<br />
</p></ol><br />
</li><li>Store the remaining resuspended parts<br />
<p> &sect; ASK: In what temperature do we keep the resuspended part?<br />
<br />
<br />
</p><br />
</h5><br />
<br />
<h1>July 7th</h1><br />
<h5><br />
<ul><br />
<li>Making 20 plates of LB Agar with Chlorampenicol added (Chloramphenicol concentration = 25 &amp;#956;g/mL</li></ul><br />
</h5><br />
<br />
<h1>July 9th</h1><br />
<h5><br />
<ul><br />
<li>Resuspending the parts we need<br />
</li><li>Transforming the plasmid into E.coli top 10 cells<br />
</li><li>Spread the transformed cells into LB Agar Plates containing appropriate antibiotics</li></ul><br />
</h5><br />
<br />
<h1>July 10th</h1><br />
<h5><br />
- No growth in positive control plate for ampicillin<br />
- Growth in negative control on chloramphenicol<br />
- No growth on RBS plate<br />
○ Possiblity: contamination<br />
- Miniprep for isolation<br />
○ 2 tubes (orange cap) @4mL broth + 4μL amp<br />
○ 5 tubes (blue c ap) @4mL broth + 2 μL chl<br />
○ 5 tubes (blue cap) @4mL broth + 2 μL IHVCB chl<br />
<br />
</h5><br />
<br />
<h1>July 11st</h1><br />
<h5><br />
<p>Today's to do list:<br />
</p><ul><br />
<li>Running electrophoresis<br />
<p> &amp;#9675; CHECK THE PROTOCOL<br />
<li>Prepare another agar plates (chloramphenicol ..., ampicilin ...)<br />
</li><li>Re-transform! <br />
<p> &amp;#9675; ASK: what's wrong with the other plates? Wrong dosage of antibiotics? Contamination?<br />
</p><p> &sect; Check the concentration of the chloramphenicol needed for each plate<br />
</p><p> &amp;#9633; Checked, 1: 2000<br />
&sect; The previous agar plates only has 1:4000 concentration of antibiotics<br />
&sect; The previous antibiotic was diluted in H2O, while there is a source which mentions that they use 25 mg/mL chloramphenicol diluted in 100% ethanol<br />
&sect; Possible reason as to why extreme growth are observed in chloramphenicol plates<br />
&amp;#9675; Prepare agar plates for 5 chloramphenicol resistant plasmids and 2 ampicillin resistant plasmids, plus positive and negative control for each antibiotic. <br />
</p><p> &sect; Plates needed: 10 + 2 + 4 + 2 = 18 plates -&gt; make 20, + 2 amp -&gt; 12:8<br />
&sect; 240mL agar for chloramphenicol, 160 mL agar for ampicilin<br />
</p></li></ul><br />
</p><p>What we did:<br />
</p><ul><br />
<li>Make another 6 plates of agar +amp and 12 plates agar +chl (+1 plate +amp for kak Gema)<br />
</li><li>Isolating the plasmids from transformed E.coli<br />
</li><li>Running the plasmid (failed)<br />
</h5><br />
<br />
<h1>July 12nd</h1><br />
<h5><br />
<p> • Electrophoresis iGEM plasmid<br />
</p><br />
</h5><br />
<br />
<h1>July 15th</h1><br />
<h5><br />
<ul><br />
<li>Re-transform the plasmid</li></ul><br />
</h5><br />
<br />
<h1>July 16th</h1><br />
<h5><br />
<ul><br />
<li>Checking the plates -&gt; contamination<br />
</li><li>Making new agar plates</li></ul><br />
</h5><br />
<br />
<h1>July 17th</h1><br />
<h5><br />
<ul><br />
<li>Primer designing<br />
</li><li>Preparing to make competent cells</li></ul><br />
</h5><br />
<br />
<h1>July 22nd</h1><br />
<h5><br />
- Re-run<br />
<br />
Well 1: marker<br />
Well 2: alfa 1/2 (colony/transform)<br />
Well 3: alfa 2/2<br />
Well 4: full 1/2<br />
Well 5: full 2/2<br />
Well 6: alfa 1/3<br />
Well 7: alfa 2/3<br />
Well 8: full 1/3<br />
Well 9: full 2/3<br />
</h5><br />
<br />
<h1>July 23rd</h1><br />
<h5><br />
<ul><br />
<li>Restricting plasmids<br />
</li><li>Running</li></ul><br />
</h5><br />
<br />
<h1>July 24th</h1><br />
<h5><br />
- Re-digest the plasmids especially containing lacZ full<br />
- Re-run<br />
- Make maxiprep<br />
<br />
<p>List of materials<br />
</p><ul><br />
<li>Digesting plasmids<br />
<ul><br />
<li>NEBuffer 10x<br />
</li><li>BSA 10x<br />
</li><li>H2O sigma (DW)<br />
</li><li>EcoRI (-800C freezer)<br />
</li><li>DNA<br />
</li></ul><br />
</li><li>Running<br />
<ul><br />
<li>Agarose<br />
</li><li>LD 6x<br />
</li><li>Marker </li></ul><br />
</li></ul><br />
</h5><br />
<br />
<h1>July 25th</h1><br />
<h5><br />
<ul><br />
<li>Large scale isolation for LacZfull<br />
</li><li>Inoculating RFP</li></ul><br />
</h5><br />
<br />
<h1>July 26th</h1><br />
<h5><br />
ul><br />
<li>Running LacZ full and lacZ alpha<br />
<br />
<br />
</li><li>Inoculating lacZ alpha<br />
</li><li>Find restriction site with buffer compatible with EcoRI XbaI SpeI and PstI</li></ul><br />
</h5><br />
<br />
<h1>July 27th</h1><br />
<h5><br />
<ul><br />
<li>Large scale isolation lacZ alpha<br />
</li><li>Making agarose<br />
</li><li>Electrophoresis<br />
<ul><br />
<li>From the UV reading of the agar, we found that the plasmid is transformed into the cell. We decided to linearize the plasmid </li></ul><br />
</li></ul><br />
</h5><br />
<br />
<h1>July 30th</h1><br />
<h5><br />
<ul><br />
<p>Labwork to do list:<br />
</p><ul><br />
<li>Digesting lacZ alpha<br />
</li><li>Running</li></ul><br />
</h5><br />
<br />
<h1>July 31st</h1><br />
<h5><br />
<ul><br />
<li>Linearizing lacZ alpha using EcoRI<br />
</li><li>Electrophoresis<br />
<ul><br />
<li>We got the correct length for lacZ alpha </li></ul><br />
</li></ul><br />
</h5><br />
<br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/ModellingTeam:UI-Indonesia/Modelling2013-10-19T02:40:43Z<p>Taufik.ha: </p>
<hr />
<div>{{:Te{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Modelling</span></h1><br />
<br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<p align="left"><br />
<h1>Activity Assay</h1><br />
</p><br />
<br><br />
<p><br />
To test the function of our split reporter, we collect the data of enzyme activity over time. We exploit the enzyme’s ability to convert <em>ortho</em><br />
-Nitrophenyl-Beta-galactoside (ONPG), a colorless substance, into ortho-nitrophenol, a visible yellow substance. We then use a spectrophotometer to<br />
quantisize the absorbance of the solution.<br />
<br><br><br />
</p><br />
<p><br />
<strong><u>Experimental setup &amp; protocol</u></strong><br />
</p><br />
<p><br />
1. The alpha and omega fragment of Beta-galactosidase was cloned into pQE-80L and pQE-81L, respectively<br />
</p><br />
<p><br />
2. The alpha and omega fragment was then expressed (in TOP10 E.coli) and purified using His tagged protein purification method<br />
</p><br />
<p><br />
3. Add an equal molar of both the alpha and omega peptide to eppendorf tube #1 - #4; alpha fragment to eppendorf tube #5 - #8; omega fragment to eppendorf<br />
tube #9 - #12; and diluted full length Beta-galactosidase to tube #13 - #16. Incubate those tubes at room temperature on an orbital rocker for 1 hour.<br />
</p><br />
<p><br />
4. At time zero, 20µL of ONPG (4mg/mL) was added into each tube<br />
</p><br />
<p><br />
5. The eppendorf tubes then incubated at room temperature for different length of time (30 min, 90 min, 3 hours, and 19 hours).<br />
</p><br />
<p><br />
6. The reaction was then terminated by adding 50µL 1M Na<sub>2</sub>CO<sub>3</sub><br />
</p><br />
<p><br />
7. The absorbance is the analysed using 420 nm light<br />
<br><br />
<br><br />
</p><br />
<p><br />
<strong><u>Result</u></strong><br />
</p><br />
<p align="center"><br />
<img<br />
src="https://static.igem.org/mediawiki/2013/6/66/Char1.JPG"><br />
<br />
<br><br />
<br><br />
</p><br />
<p><br />
<strong><u>Interpretation</u></strong><br />
</p><br />
<p><br />
The full length Beta-galactosidase reaction mix works as a positive control while both the alpha-only and omega-only reaction mix works as a negative<br />
control.<br />
</p><br />
<p><br />
The split reporter have the activity of full length Beta-galactosidase enzyme, while none of the alpha-only nor the omega-only have the enzymatic activity.<br />
This data suggests that the peptide complementation needs to occur in order to generate enzymatic activity. Previous study shows that the peptide needs<br />
many minutes to form the tetrameric structure which have the enzymatic activity. That’s why we incubate them for 1 hour after mixing the alpha and omega<br />
fragment.<br />
</p><br />
<p><br />
The data also shows that the split reporter needs longer timer to digest the same amount of ONPG compared to full length Beta-galactosidase. This data<br />
suggest that our split reporter works as expected.<br />
</p><br />
<p><br />
<br><br />
<br><br />
<h1>Stability Assay of The Split Reporter</h1><br />
</p><br />
<p><br />
We perform a stability assay to test the enzyme’s activity after freezing and storage in 4ᵒ C freezer for a different length of time.<br />
<br><br />
<br><br />
</p><br />
<p><br />
<strong><u>Experimental setup &amp; protocol</u></strong><br />
</p><br />
<p><br />
1. Add alpha and omega fragment of Beta-galactosidase to different eppendorf tube.<br />
</p><br />
<p><br />
2. Store those two tubes in 4ᵒC freezer for different length of time (2 days, 7 days, 14 days, 21 days and 28 days)<br />
</p><br />
<p><br />
3. Thaw the tube after stored in different length of time<br />
</p><br />
<p><br />
4. Add an equal molar of both the alpha and omega peptide to an eppendorf tube. Incubate those tubes at room temperature on an orbital rocker for 1 hour.<br />
</p><br />
<p><br />
5. At time zero, 20µL of ONPG (4mg/mL) was added into each tube<br />
</p><br />
<p><br />
6. The eppendorf tubes then incubated at room temperature for 3 hours<br />
</p><br />
<p><br />
7. The reaction was then terminated by adding 50µL 1M Na<sub>2</sub>CO<sub>3</sub><br />
</p><br />
<p><br />
8. The absorbance is the analysed using 420 nm light<br />
</p><br />
<p><br />
9. Results are expressed as percent signals obtained from freshly expressed enzyme after 3 hours of reaction<br />
<br><br />
<br><br />
<br />
</p><br />
<p><br />
<strong><u>Result</u></strong><br />
</p><br />
<p align="center"><br />
<img<br />
src="https://static.igem.org/mediawiki/2013/3/33/Stability_Assay.jpg"<br />
height="277"<br />
width="455"<br />
/><br />
<br />
<br><br />
<br><br />
<br />
</p><br />
<p><br />
<strong><u>Interpretation</u></strong><br />
</p><br />
<p><br />
The activity of beta-galactosidase enzyme is still above 95% after 28 days of storage. This data suggest that split reporter can be stored for a quite long<br />
time in 4ᵒC.<br />
</p><br />
<p align="left"><br />
<h1>Characterisation of Other Parts</h1><br />
</p><br />
<p><br />
Beside characterize our own part, we also characterize another old part and adding it's application. Part #BBa_J15102 and #BBa_K891003 are complementary parts that can be used to create ternary complexation mediated protein complementation based biosensor.<br />
This kind of biosensor detects a single mediator body instead of two complementing parts.<br />
</p><br />
<p><br />
<h1>Folding Time Assay</h1><br />
<strong></strong><br />
</p><br />
<p><br />
In order to create the biosensor, first we have to test the time needed for the complementing parts to form a tetramer.<br />
</p><br />
<p><br />
<strong><u>Experiment Setup and Protocol</u></strong><br />
</p><br />
<p><br />
1. Add diluted full length beta-galactosidase enzyme to eppendorf tube #1, alpha fragment to eppendorf tube #2, omega fragment to eppendorf tube #3, and<br />
mix of equimolar alpha and omega fragment to eppendorf tube #4.<br />
</p><br />
<p><br />
2. Incubate those tubes at room temperature on an orbital rocker for 1 hour.<br />
</p><br />
<p><br />
3. Add 20µL ONPG substrate to each tube<br />
</p><br />
<p><br />
4. Incubate those tubes at room temperature on an orbital locker for 30 min<br />
</p><br />
<p><br />
5. The reaction was then terminated by adding 50µL 1M Na<sub>2</sub>CO<sub>3</sub><br />
</p><br />
<p><br />
6. The absorbance is the analysed using 420 nm light<br />
</p><br />
<p><br />
<strong><u>Result</u></strong><br />
</p><br />
<p align="center"><br />
<img<br />
src="https://static.igem.org/mediawiki/2013/6/62/Folding_Time_Assay.jpg"<br />
height="468"<br />
width="470"<br />
/><br />
</p><br />
<p><br />
<strong><u>Interpretation</u></strong><br />
</p><br />
<p><br />
In the first 30 minutes of reaction, full length beta galactosidase shows higher activity; split reporter shows lower activity; alpha-only and omega-only<br />
fragment shows almost no activity. This allows us to differ full length beta-galactosidase activity from split reporter activity.<br />
</p><br />
<br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/DescriptionTeam:UI-Indonesia/Description2013-10-19T02:40:19Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Project Blue Ivy</span></h1><br />
<br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<br />
<p><h1>World’s Problem Waiting to be Solved</h1><br />
<br />
<br/><br />
Tuberculosis is still one of the worst infection-related health problem. It kills approximately 2 million people annually, making it the number one killer of single infection disease. The global epidemic is growing and becoming more dangerous. Today, tuberculosis has infected approximately a third of world’s population, even after WHO declared a global emergency on tuberculosis on 1993.<br />
<br><br />
<br>Where is the problem? Is it that hard to cure Tuberculosis? No!<br />
<br><br>WHO’s data about Tuberculosis epidemiology shows that 95% cases and 98% deaths from tuberculosis infection occur in poor countries; at the same time, 78% of world’s population dont have access to proper Tuberculosis diagnostic facilities. This clearly tells us that, the real problem in eradicating Tuberculosis is that we don’t have a reliable diagnostic tool for people in poor countries. A mathematical modelling of TB epidemiology shows that if every man and woman in this planet have an access to appropriate diagnostic facility, the prevalence of the disease will decrease 30% globally. Thus, providing a cheap, portable, reliable and easy to use TB diagnostic tool is exactly what we aim to do.<br />
<br><br />
<br>Our diagnostic tool (or biosensor) consist of an antibody fragment as the sensitive element which binds to antigen 85, a specific protein produced by Mycobacterium sp., and split Beta-galactosidase as the reporter.<br />
<br />
</p><p><h1>The Split Beta-galactosidase a.k.a The Reporter </h1><br />
Our system is based on ternary complexation mediated protein complementation principle. To put it in a simpler way, it is two proteins interacting with mediation of another protein and uses a reporter protein thatis split into two fragments, called as split reporter. Each fragment is then fused to one of the interacting protein of interest. These reporter fragment are individually inactive, and will be re-activated when the two fragments interact and re-assemble into a functioning enzyme which can convert substrate into products and gives a unique signal.<br />
<br />
<img src="https://static.igem.org/mediawiki/2013/f/f2/Desc1-r.JPG" border="0" align="middle"><br />
<br><br />
</p><p>Figure 1. Ternary complexation mediated protein complementation (Source: Stains, C.I., et al. (2010), with slight modification)<br />
<br><br />
<br><br />
We choose split Beta-Galactosidase as our reporter protein based on several considerations. First, Broome et al., (2010), have shown that they were able to split the Beta-galactosidase enzyme into two fragments that are capable to re-assemble and shows stable enzymatic activity in bacteria and in mammalian cells1. Hence, we decided to adopt their split reporter system into our biosensor. <br><br />
<br>Second, The substrate for Beta-Galactosidase, Xgal, is cheaper than the substrate for some other protein that can also functioned as split reporter, for example, luciferin (substrate for luciferase enyzme). This is important, because our goal is to make a cheap detection system to be used in suburban area.<br><br />
<br>The signal produced by Beta-Galactosidase is visible so that we dont need another device to define the infection. The signal also lasts for a long time so that we can minimize the false negative that is often occurred in short time observation. <br />
</p><p><h1>The Single Chain Fragment Variable a.k.a The Sensitive Element</h1><br />
The antibody fragment (scFv) used in our detection system is selected based on the previous experiment conducted by Ferrara et al., (2012)2. They select 111 scFvs from the previously created antibody library3 using flow cytometry. From these 111 scFvs they tested 48 of them for their specificity to Ag85, also by flow cytometry, using biotinylated Ag85; myoglobin and streptavidin-Alexa-Fluor-633 were used as negative controls. Also, they were tested to their recognition for the three different Ag85components.From the result, 14 of them were then selected for ELISA test. Three of the candidates were eliminated, leaving 11 remains. The top eight clones were further tested in an 8x8 sandwich assay, in which yeast displaying all eightselected antibodies were used as Ag85 capture reagents, and testedwith all eight scFv-Fc fusions as detection reagents. To fulfill the need of our system, we need to select two scFvs which recognize two different sites on Ag85. Based on the data on the journal, we decided to choose antibody C11 and A2 for our biosensor. <br />
<br><br />
<br />
<img src="https://static.igem.org/mediawiki/2013/0/0a/Desc2.JPG" border="0" align="middle"><br />
</p><p>Figure 2. Antibody selection method (Source:Ferrara, F., et al. (2012))<br />
<br><br>References:<br />
<br><br />
<br><br />
</p><pre width="800px";><br />
1. Broome, A.M., et al. (2010). Expanding the utility of Beta-galactosidase complementation: pieceby piece. Mol Pharm. 7(1): 60–74. doi:10.1021/mp900188e.<br />
2. Ferrara, F., et al. (2012) Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies:Application to Antigen 85, a Tuberculosis Biomarker. PLoS ONE. 7(11): e49535. doi:10.1371/journal.pone.0049535. <br />
3. Sblattero, D., Bradbury, A. (1999). Exploiting recombination in single bacteriato make large phage antibody libraries. Nature America. <br />
4. Stains, C.I., et al. (2010). A General Approach for Receptor and Antibody-TargetedDetection of Native Proteins utilizing Split-LuciferaseReassembly. ACS Chem Biol. 5(10): 943–952. doi:10.1021/cb100143m.</pre><br />
<br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/DescriptionTeam:UI-Indonesia/Description2013-10-19T02:40:00Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Project Blue Ivy</span></h1><br />
<br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<br />
<h1>World’s Problem Waiting to be Solved</h1><br />
<br />
<br/><br />
Tuberculosis is still one of the worst infection-related health problem. It kills approximately 2 million people annually, making it the number one killer of single infection disease. The global epidemic is growing and becoming more dangerous. Today, tuberculosis has infected approximately a third of world’s population, even after WHO declared a global emergency on tuberculosis on 1993.<br />
<br><br />
<br>Where is the problem? Is it that hard to cure Tuberculosis? No!<br />
<br><br>WHO’s data about Tuberculosis epidemiology shows that 95% cases and 98% deaths from tuberculosis infection occur in poor countries; at the same time, 78% of world’s population dont have access to proper Tuberculosis diagnostic facilities. This clearly tells us that, the real problem in eradicating Tuberculosis is that we don’t have a reliable diagnostic tool for people in poor countries. A mathematical modelling of TB epidemiology shows that if every man and woman in this planet have an access to appropriate diagnostic facility, the prevalence of the disease will decrease 30% globally. Thus, providing a cheap, portable, reliable and easy to use TB diagnostic tool is exactly what we aim to do.<br />
<br><br />
<br>Our diagnostic tool (or biosensor) consist of an antibody fragment as the sensitive element which binds to antigen 85, a specific protein produced by Mycobacterium sp., and split Beta-galactosidase as the reporter.<br />
<br />
</p><p><h1>The Split Beta-galactosidase a.k.a The Reporter </h1><br />
Our system is based on ternary complexation mediated protein complementation principle. To put it in a simpler way, it is two proteins interacting with mediation of another protein and uses a reporter protein thatis split into two fragments, called as split reporter. Each fragment is then fused to one of the interacting protein of interest. These reporter fragment are individually inactive, and will be re-activated when the two fragments interact and re-assemble into a functioning enzyme which can convert substrate into products and gives a unique signal.<br />
<br />
<img src="https://static.igem.org/mediawiki/2013/f/f2/Desc1-r.JPG" border="0" align="middle"><br />
<br><br />
</p><p>Figure 1. Ternary complexation mediated protein complementation (Source: Stains, C.I., et al. (2010), with slight modification)<br />
<br><br />
<br><br />
We choose split Beta-Galactosidase as our reporter protein based on several considerations. First, Broome et al., (2010), have shown that they were able to split the Beta-galactosidase enzyme into two fragments that are capable to re-assemble and shows stable enzymatic activity in bacteria and in mammalian cells1. Hence, we decided to adopt their split reporter system into our biosensor. <br><br />
<br>Second, The substrate for Beta-Galactosidase, Xgal, is cheaper than the substrate for some other protein that can also functioned as split reporter, for example, luciferin (substrate for luciferase enyzme). This is important, because our goal is to make a cheap detection system to be used in suburban area.<br><br />
<br>The signal produced by Beta-Galactosidase is visible so that we dont need another device to define the infection. The signal also lasts for a long time so that we can minimize the false negative that is often occurred in short time observation. <br />
</p><p><h1>The Single Chain Fragment Variable a.k.a The Sensitive Element</h1><br />
The antibody fragment (scFv) used in our detection system is selected based on the previous experiment conducted by Ferrara et al., (2012)2. They select 111 scFvs from the previously created antibody library3 using flow cytometry. From these 111 scFvs they tested 48 of them for their specificity to Ag85, also by flow cytometry, using biotinylated Ag85; myoglobin and streptavidin-Alexa-Fluor-633 were used as negative controls. Also, they were tested to their recognition for the three different Ag85components.From the result, 14 of them were then selected for ELISA test. Three of the candidates were eliminated, leaving 11 remains. The top eight clones were further tested in an 8x8 sandwich assay, in which yeast displaying all eightselected antibodies were used as Ag85 capture reagents, and testedwith all eight scFv-Fc fusions as detection reagents. To fulfill the need of our system, we need to select two scFvs which recognize two different sites on Ag85. Based on the data on the journal, we decided to choose antibody C11 and A2 for our biosensor. <br />
<br><br />
<br />
<img src="https://static.igem.org/mediawiki/2013/0/0a/Desc2.JPG" border="0" align="middle"><br />
</p><p>Figure 2. Antibody selection method (Source:Ferrara, F., et al. (2012))<br />
<br><br>References:<br />
<br><br />
<br><br />
</p><pre width="800px";><br />
1. Broome, A.M., et al. (2010). Expanding the utility of Beta-galactosidase complementation: pieceby piece. Mol Pharm. 7(1): 60–74. doi:10.1021/mp900188e.<br />
2. Ferrara, F., et al. (2012) Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies:Application to Antigen 85, a Tuberculosis Biomarker. PLoS ONE. 7(11): e49535. doi:10.1371/journal.pone.0049535. <br />
3. Sblattero, D., Bradbury, A. (1999). Exploiting recombination in single bacteriato make large phage antibody libraries. Nature America. <br />
4. Stains, C.I., et al. (2010). A General Approach for Receptor and Antibody-TargetedDetection of Native Proteins utilizing Split-LuciferaseReassembly. ACS Chem Biol. 5(10): 943–952. doi:10.1021/cb100143m.</pre><br />
<br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/AbstractTeam:UI-Indonesia/Abstract2013-10-19T02:39:36Z<p>Taufik.ha: </p>
<hr />
<div>{{:Te{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Abstract</span></h1><br />
<br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<p>Tuberculosis (TB) is a worldwide major health problem which infects one third of the world’s population. The absence of reliable diagnostic tool in suburban area, where TB cases are most likely found, is still a great obstacle in TB eradication effort. Seeing Indonesia as one of the high burden countries for TB, UI-Indonesia iGEM team are trying to create a reliable, portable, and easy to use diagnostic tool for detecting TB. We are constructing a biosensor consist of highly specific antibody bound to a fragment of Beta-Galactosidase as a reporter to detect the presence of protein Ag85, a novel TB biomarker. Our goal is to make a biosensor that will detect the presence of antigen 85 in blood serum of TB suspect. Positive result will be indicated with easy to detect blue color, and when it’s negative, no response will be observed. <br></p><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/DescriptionTeam:UI-Indonesia/Description2013-10-19T02:39:10Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Project Blue Ivy</span></h1><br />
<br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<br />
<br />
<p><h1>World’s Problem Waiting to be Solved</h1><br />
<br />
<br/><br />
Tuberculosis is still one of the worst infection-related health problem. It kills approximately 2 million people annually, making it the number one killer of single infection disease. The global epidemic is growing and becoming more dangerous. Today, tuberculosis has infected approximately a third of world’s population, even after WHO declared a global emergency on tuberculosis on 1993.<br />
<br><br />
<br>Where is the problem? Is it that hard to cure Tuberculosis? No!<br />
<br><br>WHO’s data about Tuberculosis epidemiology shows that 95% cases and 98% deaths from tuberculosis infection occur in poor countries; at the same time, 78% of world’s population dont have access to proper Tuberculosis diagnostic facilities. This clearly tells us that, the real problem in eradicating Tuberculosis is that we don’t have a reliable diagnostic tool for people in poor countries. A mathematical modelling of TB epidemiology shows that if every man and woman in this planet have an access to appropriate diagnostic facility, the prevalence of the disease will decrease 30% globally. Thus, providing a cheap, portable, reliable and easy to use TB diagnostic tool is exactly what we aim to do.<br />
<br><br />
<br>Our diagnostic tool (or biosensor) consist of an antibody fragment as the sensitive element which binds to antigen 85, a specific protein produced by Mycobacterium sp., and split Beta-galactosidase as the reporter.<br />
<br />
</p><p><h1>The Split Beta-galactosidase a.k.a The Reporter </h1><br />
Our system is based on ternary complexation mediated protein complementation principle. To put it in a simpler way, it is two proteins interacting with mediation of another protein and uses a reporter protein thatis split into two fragments, called as split reporter. Each fragment is then fused to one of the interacting protein of interest. These reporter fragment are individually inactive, and will be re-activated when the two fragments interact and re-assemble into a functioning enzyme which can convert substrate into products and gives a unique signal.<br />
<br />
<img src="https://static.igem.org/mediawiki/2013/f/f2/Desc1-r.JPG" border="0" align="middle"><br />
<br><br />
</p><p>Figure 1. Ternary complexation mediated protein complementation (Source: Stains, C.I., et al. (2010), with slight modification)<br />
<br><br />
<br><br />
We choose split Beta-Galactosidase as our reporter protein based on several considerations. First, Broome et al., (2010), have shown that they were able to split the Beta-galactosidase enzyme into two fragments that are capable to re-assemble and shows stable enzymatic activity in bacteria and in mammalian cells1. Hence, we decided to adopt their split reporter system into our biosensor. <br><br />
<br>Second, The substrate for Beta-Galactosidase, Xgal, is cheaper than the substrate for some other protein that can also functioned as split reporter, for example, luciferin (substrate for luciferase enyzme). This is important, because our goal is to make a cheap detection system to be used in suburban area.<br><br />
<br>The signal produced by Beta-Galactosidase is visible so that we dont need another device to define the infection. The signal also lasts for a long time so that we can minimize the false negative that is often occurred in short time observation. <br />
</p><p><h1>The Single Chain Fragment Variable a.k.a The Sensitive Element</h1><br />
The antibody fragment (scFv) used in our detection system is selected based on the previous experiment conducted by Ferrara et al., (2012)2. They select 111 scFvs from the previously created antibody library3 using flow cytometry. From these 111 scFvs they tested 48 of them for their specificity to Ag85, also by flow cytometry, using biotinylated Ag85; myoglobin and streptavidin-Alexa-Fluor-633 were used as negative controls. Also, they were tested to their recognition for the three different Ag85components.From the result, 14 of them were then selected for ELISA test. Three of the candidates were eliminated, leaving 11 remains. The top eight clones were further tested in an 8x8 sandwich assay, in which yeast displaying all eightselected antibodies were used as Ag85 capture reagents, and testedwith all eight scFv-Fc fusions as detection reagents. To fulfill the need of our system, we need to select two scFvs which recognize two different sites on Ag85. Based on the data on the journal, we decided to choose antibody C11 and A2 for our biosensor. <br />
<br><br />
<br />
<img src="https://static.igem.org/mediawiki/2013/0/0a/Desc2.JPG" border="0" align="middle"><br />
</p><p>Figure 2. Antibody selection method (Source:Ferrara, F., et al. (2012))<br />
<br><br>References:<br />
<br><br />
<br><br />
</p><pre width="800px";><br />
1. Broome, A.M., et al. (2010). Expanding the utility of Beta-galactosidase complementation: pieceby piece. Mol Pharm. 7(1): 60–74. doi:10.1021/mp900188e.<br />
2. Ferrara, F., et al. (2012) Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies:Application to Antigen 85, a Tuberculosis Biomarker. PLoS ONE. 7(11): e49535. doi:10.1371/journal.pone.0049535. <br />
3. Sblattero, D., Bradbury, A. (1999). Exploiting recombination in single bacteriato make large phage antibody libraries. Nature America. <br />
4. Stains, C.I., et al. (2010). A General Approach for Receptor and Antibody-TargetedDetection of Native Proteins utilizing Split-LuciferaseReassembly. ACS Chem Biol. 5(10): 943–952. doi:10.1021/cb100143m.</pre><br />
<br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/TeamTeam:UI-Indonesia/Team2013-10-19T02:38:53Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:15px 15px;"><br />
<h1>The Great Team</h1><br />
<br />
<p>Our project is totally an undergraduate project. All the works in our group are done by undergraduate, starting from brainstorming the idea, wet work on the bench, lab troubleshooting, constructing wiki as well as sponsors and donors hunting, It has been a long project but it is totally rewarding. We have fun and learn a lot of things. All of team members have increase the lab work skills, journal reading skills as well as negotiating skills.</p><br />
<br />
Team Description:<br />
<br />
<br />
<table style="margin-left:120px; margin-right:120px; background-color:transparent"><br />
<br />
<tr><br />
<td><h2>Supervisors</h2><img src="https://static.igem.org/mediawiki/2013/6/6a/Dr_Budi.jpg" align="center"></td><br />
<td><h3>Dr. dr. Budiman Bela, Sp.MK (K)</h3><br />
<p>Head of Clinical Microbiology laboratory at Faculty of Medicine, Universitas Indonesia.</p></td><br />
</tr><br />
<br />
<tr><br />
<td><img src="https://static.igem.org/mediawiki/2013/6/69/Pa_Abin.jpg" align="center"></td><br />
<td><h3>Drs. Abinawanto, M.Si</h3><br />
<p>Head of Genetics laboratory at Faculty of Mathematics and Natural Sciences, Universitas Indonesia.</p></td><br />
</tr><br />
<br />
<br />
<tr><br />
<td><h2>Students</h2><br />
<img src="https://static.igem.org/mediawiki/2013/c/c3/Hanifi.jpg" align="center"></td><br />
<td><h4>Muhammad Hanifi (Medicine, ’11)</h4><p>The leader of the group. Sing along to each and every song on the radio while we’re working in the lab. Even though sometimes he does not know the lyrics. Or the melody. Or both.</p></td><br />
</tr><br />
<br />
<tr><br />
<td><br />
<img src="https://static.igem.org/mediawiki/2013/f/f7/Danny.jpg"></td><br />
<td><h4>Achmad Danny Gazali (Biology, ’10)</h4><p>The movie maker. Records and takes pics of our activities. Has the Yao Ming-meme-face when laughing.</p><br />
</td><br />
</tr> <br />
<br />
<tr><br />
<td><br />
<img src="https://static.igem.org/mediawiki/2013/c/ce/Wian.jpg"></td><br />
<td><h4>Dwiantari Satyapertiwi (Chemical Engineering, ’11)</h4><p>The maker of this page.The other members don’t know what I’m writing, so, the information provided here may or may not be accurate ;)</p></td><br />
</tr><br />
<br />
<tr><br />
<td><br />
<img src="https://static.igem.org/mediawiki/2013/8/86/Teguh.jpg"></td><br />
<td><h4>Mohamad Teguh Gumelar (Bioprocess Technology, ’11)</h4><p>The naïve one. Constantly got tricked by the other members. Takes lots, yes, LOTS of selfies.</p></td><br />
</tr><br />
<br />
<tr> <br />
<td><br />
<img src="https://static.igem.org/mediawiki/2013/b/b6/Widia.jpg"></td><br />
<td><h4>Putu Ayuwidia Ekaputri (Medicine, ’10)</h4><p>The random Sanguine. Takes bus alone to her birthplace (approximately 3 hours from her current address), on her birthday, just to take picture of herself in front of the hospital she was born in. And it’s unplanned.<p></td><br />
</tr><br />
<br />
</tr><br />
<td><br />
<img src="https://static.igem.org/mediawiki/2013/b/b5/Rahdi.jpg"></td><br />
<td><h4>Rahdi Dewin Marzaini (Medicine, ’11)</h4><p>The linguist. Or maybe language maniac. Knows maybe 10 languages apart from his mother tongue. Found his soulmate in High School, who is no other than...Achmad Danny Gazali.</p></td><br />
</tr><br />
<br />
<tr><br />
<td><br />
<img src="https://static.igem.org/mediawiki/2013/e/ed/Taufik.jpg"></td><br />
<td><h4>Taufik Hidayat Abdullah (Bioprocess Technology, ’11)</h4><p>The web designer. Also the main personnel in the lab. The one who has the best “bench-flip” expression.</p></td><br />
</tr><br />
<br />
<tr><br />
<td><br />
<img src="https://static.igem.org/mediawiki/2013/e/ec/Wisnu.jpg"></td><br />
<td><h4>Wisnu Tafroji (Biology, ’10)</h4><p>The one who is worried about ripping more than two pairs of gloves everytime he enters the lab.</p></td><br />
</tr><br />
<br />
</table> <br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/AbstractTeam:UI-Indonesia/Abstract2013-10-19T02:37:38Z<p>Taufik.ha: </p>
<hr />
<div>{{:Te{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Abstract</span></h1><br />
<br />
<div style="background-color:white ;border:2px solid;border-radius:15px; padding:10px 10px;"><br />
<p>Tuberculosis (TB) is a worldwide major health problem which infects one third of the world’s population. The absence of reliable diagnostic tool in suburban area, where TB cases are most likely found, is still a great obstacle in TB eradication effort. Seeing Indonesia as one of the high burden countries for TB, UI-Indonesia iGEM team are trying to create a reliable, portable, and easy to use diagnostic tool for detecting TB. We are constructing a biosensor consist of highly specific antibody bound to a fragment of Beta-Galactosidase as a reporter to detect the presence of protein Ag85, a novel TB biomarker. Our goal is to make a biosensor that will detect the presence of antigen 85 in blood serum of TB suspect. Positive result will be indicated with easy to detect blue color, and when it’s negative, no response will be observed. <br></p><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/AbstractTeam:UI-Indonesia/Abstract2013-10-19T02:17:38Z<p>Taufik.ha: </p>
<hr />
<div>{{:Te{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Abstract</span></h1><br />
<br />
<div style="background-color:white ;border:2px solid;<br />
border-radius:15px;"><br />
<p><br>Tuberculosis (TB) is a worldwide major health problem which infects one third of the world’s population. The absence of reliable diagnostic tool in suburban area, where TB cases are most likely found, is still a great obstacle in TB eradication effort. Seeing Indonesia as one of the high burden countries for TB, UI-Indonesia iGEM team are trying to create a reliable, portable, and easy to use diagnostic tool for detecting TB. We are constructing a biosensor consist of highly specific antibody bound to a fragment of Beta-Galactosidase as a reporter to detect the presence of protein Ag85, a novel TB biomarker. Our goal is to make a biosensor that will detect the presence of antigen 85 in blood serum of TB suspect. Positive result will be indicated with easy to detect blue color, and when it’s negative, no response will be observed. <br></p><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/SafetyTeam:UI-Indonesia/Safety2013-10-19T01:58:58Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Safety Form</span></h1><br />
<br />
<div style="background-color:white";><br />
<p><img src="https://static.igem.org/mediawiki/2013/7/77/UiSafety1.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/9/9e/Safety2.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/2/23/UiSafety3.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/b/be/Safety4.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/3/30/Safety5.JPG.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/4/45/UiSafety6.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/3/35/Safety7.JPG" border="0" align="middle"></p><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/SafetyTeam:UI-Indonesia/Safety2013-10-19T01:58:33Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>Safety Form</span></h1><br />
<br />
<div style="background-color:white";><br />
<p><img src="https://static.igem.org/mediawiki/2013/7/77/UiSafety1.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/9/9e/Safety2.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/2/23/UiSafety3.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/b/be/Safety4.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/3/30/Safety5.JPG.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/4/45/UiSafety6.JPG" border="0" align="middle"></p><br />
<p><img src="https://static.igem.org/mediawiki/2013/7/77/UiSafety7.JPG" border="0" align="middle"></p><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/headerTeam:UI-Indonesia/header2013-10-19T01:56:40Z<p>Taufik.ha: </p>
<hr />
<div><!-- *** Mencoba buat template isinya header, menu, background *** --><br />
<html><br />
<style><br />
body{position:absolute; top:0px; width:100%; height:100%;}<br />
<br />
#contentSub {<br />
display:none;<br />
}<br />
body {<br />
background: #fff url('https://static.igem.org/mediawiki/2013/c/c1/Bg3ui.jpg') no-repeat fixed;<br />
background-size: 1350px ;<br />
background-position: center top;<br />
}<br />
<br />
#top-section{<br />
height:0px;<br />
border:none;<br />
width:980px;<br />
margin:0 auto;<br />
padding:0 0 0 0;<br />
background-color:transparent;<br />
overflow:hide;<br />
<br />
}<br />
#p-logo{display:none;}<br />
#search-controls{display:none;}<br />
#top{display:none;}<br />
.firstHeading{display:none;}<br />
#footer-box{display:none;}<br />
#catlinks{display:none;}<br />
#content {background-color:transparent; border-left-width:0px; border-right-width:0px; padding: 0px 5px 0px 5px}<br />
</style><br />
<body><br />
<!-- header of the page --><br />
<header><br />
<div id= "header" style= "float: none; width: 960px;<br />
height: auto;"><br />
<br />
<!-- *** Mulai kode gambar header oleh Teguh *** --><br />
<img src="https://static.igem.org/mediawiki/2013/6/67/Header_New_Design.png" border="0"><br />
<br />
<br />
<!-- *** Akhir kode gambar header oleh Teguh *** --><br />
<br />
</header><br />
<div><br />
<br />
<!-- *** Menu Style CSS From MIT 2012, Thankyou..!! :D *** --><br />
<style><br />
.nav{<br />
float:left;<br />
width:100%;<br />
margin:0;<br />
list-style:none;<br />
position:relative;<br />
z-index:5;<br />
<br />
/*CHANGE THIS TO CENTER*/<br />
padding:0 185px; /*12px;*/<br />
}<br />
<br />
.nav li{<br />
float:left;<br />
position:relative;<br />
display:inline;<br />
line-height: 2em;<br />
}<br />
<br />
.nav a{<br />
float:left;<br />
height:2em;<br />
padding: 0 1.3em;<br />
color:white;<br />
/*Padding: bottom, right, top, left */<br />
/*padding:0.2em 1.3em 0.2em 1.3em;*/<br />
border-right:1px solid white;<br />
white-space:nowrap;<br />
}<br />
<br />
.nav a:hover{<br />
color:#00FFFF !important;<br />
}<br />
<br />
/*--- DROPDOWN ---*/<br />
.nav ul{<br />
background:#fff; <br />
list-style:none;<br />
position:absolute;<br />
left:-9999px; /* Hide off-screen when not needed (this is more accessible than display:none;) */<br />
margin-top: 25px;<br />
margin-left:0;<br />
margin-right:0;<br />
<br />
}<br />
<br />
<br />
.nav ul li{<br />
padding-top:1px;<br />
}<br />
<br />
.nav ul a{<br />
padding-top: 3px;<br />
padding-bottom:3px; /*Introducing a padding between the li and the a give the illusion spaced items */<br />
white-space:normal;<br />
text-decoration:none;<br />
height:auto;<br />
width: 13em;<br />
line-height: 1.5em;<br />
}<br />
<br />
.nav li:hover ul{ /* Display the dropdown on hover */<br />
left:0; /* Bring back on-screen when needed */<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover a{ /* These create persistent hover states, meaning the top-most link stays 'hovered' even when your cursor has moved down the list. */<br />
<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul a{ /* The persistent hover state does however create a global style for links even before they're hovered. Here we undo these effects. */<br />
background:#121212;<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul li a:hover{ /* Here we define the most explicit hover states--what happens when you hover each individual link. */<br />
background:#8a8a88;<br />
text-decoration:none;<br />
}<br />
</style><br />
<!-- *** End of Menubar CSS style code from MIT, Thankyouuu :D *** --><br />
<br />
<div id="border-top"></div> <br />
<br />
<!-- *** Percobaan kode menu dropdown MIT *** --><br />
<ul class="nav"><br />
<br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia">HOME</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Team">TEAM</a><br />
<!-- ***<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Member">Members</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Lab">Our Lab</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Gallery">Gallery</a></li><br />
</ul>*** --><br />
</li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Description">PROJECT</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Description">Overview</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Abstract">Abstract</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Parts">Parts</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Modelling">Modelling</a></li><br />
<!-- ***<li><a href="https://2013.igem.org/Team:UI-Indonesia/JudgingForm">Judging Form</a></li>*** --><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/July">NOTEBOOK</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/July">July</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/August">August</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/September">September</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices">HUMAN PRACTICES</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
</ul><br />
<hr color:"white";> <br />
</html></div>Taufik.hahttp://2013.igem.org/File:Pcromega.jpgFile:Pcromega.jpg2013-09-30T19:59:15Z<p>Taufik.ha: </p>
<hr />
<div></div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/JulyTeam:UI-Indonesia/July2013-09-28T00:35:33Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>July</span></h1><br />
<div style="background-color:white";><br />
<br />
<h1>July 1st </h1><br />
<h5><br />
<p>What to do:<br />
</p><ul><br />
<li>Resuspend plasmids from the iGEM kit<br />
</li><li>Transform to E.coli cell<br />
</li><li>Store the remaining suspended plasmid<br />
<br />
</li></ul><br />
<p>Reagents and equipments<br />
</p><ul><br />
<li>Reagents<br />
<ol><br />
<li>Distilled water<br />
</li><li>Ice<br />
</li><li>50 µL competent cells for parts of choice<br />
</li><li>50 µL competent cells for positive control<br />
</li><li>Positive control<br />
</li><li>SOC Media<br />
</li><li>Agar plates with appropriate antibiotic<br />
<p> &sect; NOTE: this needs to be prepared beforehand. <br />
</p></ol><br />
</li><li>Equipments<br />
<ol><br />
<li>10 and 200 µL pipettes<br />
</li><li>Timer<br />
</li><li>Water bath (420C)<br />
<br />
<dl><br />
<dt>Procedures</dt><br />
<dd> </dd></dl><br />
</li></ol><br />
</li><li>Resuspending the part<br />
<ol><br />
<li>Mark the location of the parts using a pen or marker<br />
</li><li>Punch a hole through the foil using a pipette tip<br />
</li><li>Add 10µL of dH2O<br />
</li><li>Mix thoroughly by aspirating up and down a few times<br />
</li></ol><br />
</li><li>Inoculating the DNA<br />
<ol><br />
<li>Add 1µL of the suspended DNA to the competent cells<br />
<p> &sect; The competent cells must be kept on ice before and after the addition of the suspended DNA<br />
&sect; Make sure to mark the lid of the eppendorf tube, which tube is filled with C.C +DNA and which tube is filled with positive control.<br />
<li>Add positive control to the competent cells<br />
<p> &sect; ASK: What strain of E.coli are we going to use as the competent cell? E.coli codon plus? Top10? BL21? pLysS?<br />
<br />
</p></ol><br />
</li><li>Incubate on ice for 30 minutes<br />
<br />
</li><li>Heatshock in water bath for 1 minute<br />
<br />
</li><li>Incubate on ice for 5 min<br />
<br />
</li><li>Add 200 µL of SOC media to each transformation<br />
<p> &sect; ASK: Is it available in IHVCB? If SOC media is not available, can we replace it with regular LB broth? <br />
<br />
<li>Incubate for 2 hours at 370C<br />
<br />
</li><li>Plate out the tranformation<br />
<ol><br />
<li>Plate 20 µL to one agar plate, and the remainder to the separate plate.<br />
</li><li>Use beads to spread evenly<br />
</li><li>Incubate overnight (16hrs) at 370C<br />
<p> &sect; ASK: What antibiotics do we need? Is it available in IHVCB? <br />
<br />
</p></ol><br />
</li><li>Store the remaining resuspended parts<br />
<p> &sect; ASK: In what temperature do we keep the resuspended part?<br />
<br />
<br />
</p><br />
</h5><br />
<br />
<h1>July 7th</h1><br />
<h5><br />
<ul><br />
<li>Making 20 plates of LB Agar with Chlorampenicol added (Chloramphenicol concentration = 25 &amp;#956;g/mL</li></ul><br />
</h5><br />
<br />
<h1>July 9th</h1><br />
<h5><br />
<ul><br />
<li>Resuspending the parts we need<br />
</li><li>Transforming the plasmid into E.coli top 10 cells<br />
</li><li>Spread the transformed cells into LB Agar Plates containing appropriate antibiotics</li></ul><br />
</h5><br />
<br />
<h1>July 10th</h1><br />
<h5><br />
- No growth in positive control plate for ampicillin<br />
- Growth in negative control on chloramphenicol<br />
- No growth on RBS plate<br />
○ Possiblity: contamination<br />
- Miniprep for isolation<br />
○ 2 tubes (orange cap) @4mL broth + 4μL amp<br />
○ 5 tubes (blue c ap) @4mL broth + 2 μL chl<br />
○ 5 tubes (blue cap) @4mL broth + 2 μL IHVCB chl<br />
<br />
</h5><br />
<br />
<h1>July 11st</h1><br />
<h5><br />
<p>Today's to do list:<br />
</p><ul><br />
<li>Running electrophoresis<br />
<p> &amp;#9675; CHECK THE PROTOCOL<br />
<li>Prepare another agar plates (chloramphenicol ..., ampicilin ...)<br />
</li><li>Re-transform! <br />
<p> &amp;#9675; ASK: what's wrong with the other plates? Wrong dosage of antibiotics? Contamination?<br />
</p><p> &sect; Check the concentration of the chloramphenicol needed for each plate<br />
</p><p> &amp;#9633; Checked, 1: 2000<br />
&sect; The previous agar plates only has 1:4000 concentration of antibiotics<br />
&sect; The previous antibiotic was diluted in H2O, while there is a source which mentions that they use 25 mg/mL chloramphenicol diluted in 100% ethanol<br />
&sect; Possible reason as to why extreme growth are observed in chloramphenicol plates<br />
&amp;#9675; Prepare agar plates for 5 chloramphenicol resistant plasmids and 2 ampicillin resistant plasmids, plus positive and negative control for each antibiotic. <br />
</p><p> &sect; Plates needed: 10 + 2 + 4 + 2 = 18 plates -&gt; make 20, + 2 amp -&gt; 12:8<br />
&sect; 240mL agar for chloramphenicol, 160 mL agar for ampicilin<br />
</p></li></ul><br />
</p><p>What we did:<br />
</p><ul><br />
<li>Make another 6 plates of agar +amp and 12 plates agar +chl (+1 plate +amp for kak Gema)<br />
</li><li>Isolating the plasmids from transformed E.coli<br />
</li><li>Running the plasmid (failed)<br />
</h5><br />
<br />
<h1>July 12nd</h1><br />
<h5><br />
<p> • Electrophoresis iGEM plasmid<br />
</p><br />
</h5><br />
<br />
<h1>July 15th</h1><br />
<h5><br />
<ul><br />
<li>Re-transform the plasmid</li></ul><br />
</h5><br />
<br />
<h1>July 16th</h1><br />
<h5><br />
<ul><br />
<li>Checking the plates -&gt; contamination<br />
</li><li>Making new agar plates</li></ul><br />
</h5><br />
<br />
<h1>July 17th</h1><br />
<h5><br />
<ul><br />
<li>Primer designing<br />
</li><li>Preparing to make competent cells</li></ul><br />
</h5><br />
<br />
<h1>July 22nd</h1><br />
<h5><br />
- Re-run<br />
<br />
Well 1: marker<br />
Well 2: alfa 1/2 (colony/transform)<br />
Well 3: alfa 2/2<br />
Well 4: full 1/2<br />
Well 5: full 2/2<br />
Well 6: alfa 1/3<br />
Well 7: alfa 2/3<br />
Well 8: full 1/3<br />
Well 9: full 2/3<br />
</h5><br />
<br />
<h1>July 23rd</h1><br />
<h5><br />
<ul><br />
<li>Restricting plasmids<br />
</li><li>Running</li></ul><br />
</h5><br />
<br />
<h1>July 24th</h1><br />
<h5><br />
- Re-digest the plasmids especially containing lacZ full<br />
- Re-run<br />
- Make maxiprep<br />
<br />
<p>List of materials<br />
</p><ul><br />
<li>Digesting plasmids<br />
<ul><br />
<li>NEBuffer 10x<br />
</li><li>BSA 10x<br />
</li><li>H2O sigma (DW)<br />
</li><li>EcoRI (-800C freezer)<br />
</li><li>DNA<br />
</li></ul><br />
</li><li>Running<br />
<ul><br />
<li>Agarose<br />
</li><li>LD 6x<br />
</li><li>Marker </li></ul><br />
</li></ul><br />
</h5><br />
<br />
<h1>July 25th</h1><br />
<h5><br />
<ul><br />
<li>Large scale isolation for LacZfull<br />
</li><li>Inoculating RFP</li></ul><br />
</h5><br />
<br />
<h1>July 26th</h1><br />
<h5><br />
ul><br />
<li>Running LacZ full and lacZ alpha<br />
<br />
<br />
</li><li>Inoculating lacZ alpha<br />
</li><li>Find restriction site with buffer compatible with EcoRI XbaI SpeI and PstI</li></ul><br />
</h5><br />
<br />
<h1>July 27th</h1><br />
<h5><br />
<ul><br />
<li>Large scale isolation lacZ alpha<br />
</li><li>Making agarose<br />
</li><li>Electrophoresis<br />
<ul><br />
<li>From the UV reading of the agar, we found that the plasmid is transformed into the cell. We decided to linearize the plasmid </li></ul><br />
</li></ul><br />
</h5><br />
<br />
<h1>July 30th</h1><br />
<h5><br />
<ul><br />
<p>Labwork to do list:<br />
</p><ul><br />
<li>Digesting lacZ alpha<br />
</li><li>Running</li></ul><br />
</h5><br />
<br />
<h1>July 31st</h1><br />
<h5><br />
<ul><br />
<li>Linearizing lacZ alpha using EcoRI<br />
</li><li>Electrophoresis<br />
<ul><br />
<li>We got the correct length for lacZ alpha </li></ul><br />
</li></ul><br />
</h5><br />
<br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/JulyTeam:UI-Indonesia/July2013-09-28T00:02:50Z<p>Taufik.ha: Created page with "{{:Team:UI-Indonesia/header}} <html> <br> <br> <br> <h1><span id= "The_Great_Team"; style="color:white";>July</span></h1> <div style="background-color:white";> <h1>July 1s..."</p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>July</span></h1><br />
<div style="background-color:white";><br />
<br />
<h1>July 1st </h1><br />
<h5><br />
<p>What to do:<br />
</p><ul><br />
<li>Resuspend plasmids from the iGEM kit<br />
</li><li>Transform to E.coli cell<br />
</li><li>Store the remaining suspended plasmid<br />
<br />
</li></ul><br />
<p>Reagents and equipments<br />
</p><ul><br />
<li>Reagents<br />
<ol><br />
<li>Distilled water<br />
</li><li>Ice<br />
</li><li>50 µL competent cells for parts of choice<br />
</li><li>50 µL competent cells for positive control<br />
</li><li>Positive control<br />
</li><li>SOC Media<br />
</li><li>Agar plates with appropriate antibiotic<br />
<p> &sect; NOTE: this needs to be prepared beforehand. <br />
</p></ol><br />
</li><li>Equipments<br />
<ol><br />
<li>10 and 200 µL pipettes<br />
</li><li>Timer<br />
</li><li>Water bath (420C)<br />
<br />
<dl><br />
<dt>Procedures</dt><br />
<dd> </dd></dl><br />
</li></ol><br />
</li><li>Resuspending the part<br />
<ol><br />
<li>Mark the location of the parts using a pen or marker<br />
</li><li>Punch a hole through the foil using a pipette tip<br />
</li><li>Add 10µL of dH2O<br />
</li><li>Mix thoroughly by aspirating up and down a few times<br />
</li></ol><br />
</li><li>Inoculating the DNA<br />
<ol><br />
<li>Add 1µL of the suspended DNA to the competent cells<br />
<p> &sect; The competent cells must be kept on ice before and after the addition of the suspended DNA<br />
&sect; Make sure to mark the lid of the eppendorf tube, which tube is filled with C.C +DNA and which tube is filled with positive control.<br />
<li>Add positive control to the competent cells<br />
<p> &sect; ASK: What strain of E.coli are we going to use as the competent cell? E.coli codon plus? Top10? BL21? pLysS?<br />
<br />
</p></ol><br />
</li><li>Incubate on ice for 30 minutes<br />
<br />
</li><li>Heatshock in water bath for 1 minute<br />
<br />
</li><li>Incubate on ice for 5 min<br />
<br />
</li><li>Add 200 µL of SOC media to each transformation<br />
<p> &sect; ASK: Is it available in IHVCB? If SOC media is not available, can we replace it with regular LB broth? <br />
<br />
<li>Incubate for 2 hours at 370C<br />
<br />
</li><li>Plate out the tranformation<br />
<ol><br />
<li>Plate 20 µL to one agar plate, and the remainder to the separate plate.<br />
</li><li>Use beads to spread evenly<br />
</li><li>Incubate overnight (16hrs) at 370C<br />
<p> &sect; ASK: What antibiotics do we need? Is it available in IHVCB? <br />
<br />
</p></ol><br />
</li><li>Store the remaining resuspended parts<br />
<p> &sect; ASK: In what temperature do we keep the resuspended part?<br />
<br />
<br />
</p><br />
</h5><br />
<br />
<h1>July 7th</h1><br />
<h5><br />
<ul><br />
<li>Making 20 plates of LB Agar with Chlorampenicol added (Chloramphenicol concentration = 25 &amp;#956;g/mL</li></ul><br />
</h5><br />
<br />
<h1>July 9th</h1><br />
<h5><br />
<ul><br />
<li>Resuspending the parts we need<br />
</li><li>Transforming the plasmid into E.coli top 10 cells<br />
</li><li>Spread the transformed cells into LB Agar Plates containing appropriate antibiotics</li></ul><br />
</h5><br />
<br />
<h1>July 10th</h1><br />
<h5><br />
- No growth in positive control plate for ampicillin<br />
- Growth in negative control on chloramphenicol<br />
- No growth on RBS plate<br />
○ Possiblity: contamination<br />
- Miniprep for isolation<br />
○ 2 tubes (orange cap) @4mL broth + 4μL amp<br />
○ 5 tubes (blue c ap) @4mL broth + 2 μL chl<br />
○ 5 tubes (blue cap) @4mL broth + 2 μL IHVCB chl<br />
<br />
</h5><br />
<br />
<h1>July 11st</h1><br />
<h5><br />
<p>Today's to do list:<br />
</p><ul><br />
<li>Running electrophoresis<br />
<p> &amp;#9675; CHECK THE PROTOCOL<br />
<li>Prepare another agar plates (chloramphenicol ..., ampicilin ...)<br />
</li><li>Re-transform! <br />
<p> &amp;#9675; ASK: what's wrong with the other plates? Wrong dosage of antibiotics? Contamination?<br />
</p><p> &sect; Check the concentration of the chloramphenicol needed for each plate<br />
</p><p> &amp;#9633; Checked, 1: 2000<br />
&sect; The previous agar plates only has 1:4000 concentration of antibiotics<br />
&sect; The previous antibiotic was diluted in H2O, while there is a source which mentions that they use 25 mg/mL chloramphenicol diluted in 100% ethanol<br />
&sect; Possible reason as to why extreme growth are observed in chloramphenicol plates<br />
&amp;#9675; Prepare agar plates for 5 chloramphenicol resistant plasmids and 2 ampicillin resistant plasmids, plus positive and negative control for each antibiotic. <br />
</p><p> &sect; Plates needed: 10 + 2 + 4 + 2 = 18 plates -&gt; make 20, + 2 amp -&gt; 12:8<br />
&sect; 240mL agar for chloramphenicol, 160 mL agar for ampicilin<br />
</p></li></ul><br />
</p><p>What we did:<br />
</p><ul><br />
<li>Make another 6 plates of agar +amp and 12 plates agar +chl (+1 plate +amp for kak Gema)<br />
</li><li>Isolating the plasmids from transformed E.coli<br />
</li><li>Running the plasmid (failed)<br />
</h5><br />
<br />
<h1>July 12nd</h1><br />
<h5><br />
<p> • Electrophoresis iGEM plasmid<br />
</p><br />
</h5><br />
<br />
<br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/headerTeam:UI-Indonesia/header2013-09-27T23:33:29Z<p>Taufik.ha: </p>
<hr />
<div><!-- *** Mencoba buat template isinya header, menu, background *** --><br />
<html><br />
<style><br />
body{position:absolute; top:0px; width:100%; height:100%;}<br />
<br />
#contentSub {<br />
display:none;<br />
}<br />
body {<br />
background: #fff url('https://static.igem.org/mediawiki/2013/c/c1/Bg3ui.jpg') no-repeat fixed;<br />
background-size: 1350px ;<br />
background-position: center top;<br />
}<br />
<br />
#top-section{<br />
height:0px;<br />
border:none;<br />
width:980px;<br />
margin:0 auto;<br />
padding:0 0 0 0;<br />
background-color:transparent;<br />
overflow:hide;<br />
<br />
}<br />
#p-logo{display:none;}<br />
#search-controls{display:none;}<br />
#top{display:none;}<br />
.firstHeading{display:none;}<br />
#footer-box{display:none;}<br />
#catlinks{display:none;}<br />
#content {background-color:transparent; border-left-width:0px; border-right-width:0px; padding: 0px 5px 0px 5px}<br />
</style><br />
<body><br />
<!-- header of the page --><br />
<header><br />
<div id= "header" style= "float: none; width: 960px;<br />
height: auto;"><br />
<br />
<!-- *** Mulai kode gambar header oleh Teguh *** --><br />
<img src="https://static.igem.org/mediawiki/2013/6/67/Header_New_Design.png" border="0"><br />
<br />
<br />
<!-- *** Akhir kode gambar header oleh Teguh *** --><br />
<br />
</header><br />
<div><br />
<br />
<!-- *** Menu Style CSS From MIT 2012, Thankyou..!! :D *** --><br />
<style><br />
.nav{<br />
float:left;<br />
width:100%;<br />
margin:0;<br />
list-style:none;<br />
position:relative;<br />
z-index:5;<br />
<br />
/*CHANGE THIS TO CENTER*/<br />
padding:0 185px; /*12px;*/<br />
}<br />
<br />
.nav li{<br />
float:left;<br />
position:relative;<br />
display:inline;<br />
line-height: 2em;<br />
}<br />
<br />
.nav a{<br />
float:left;<br />
height:2em;<br />
padding: 0 1.3em;<br />
color:white;<br />
/*Padding: bottom, right, top, left */<br />
/*padding:0.2em 1.3em 0.2em 1.3em;*/<br />
border-right:1px solid white;<br />
white-space:nowrap;<br />
}<br />
<br />
.nav a:hover{<br />
color:#00FFFF !important;<br />
}<br />
<br />
/*--- DROPDOWN ---*/<br />
.nav ul{<br />
background:#fff; <br />
list-style:none;<br />
position:absolute;<br />
left:-9999px; /* Hide off-screen when not needed (this is more accessible than display:none;) */<br />
margin-top: 25px;<br />
margin-left:0;<br />
margin-right:0;<br />
<br />
}<br />
<br />
<br />
.nav ul li{<br />
padding-top:1px;<br />
}<br />
<br />
.nav ul a{<br />
padding-top: 3px;<br />
padding-bottom:3px; /*Introducing a padding between the li and the a give the illusion spaced items */<br />
white-space:normal;<br />
text-decoration:none;<br />
height:auto;<br />
width: 13em;<br />
line-height: 1.5em;<br />
}<br />
<br />
.nav li:hover ul{ /* Display the dropdown on hover */<br />
left:0; /* Bring back on-screen when needed */<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover a{ /* These create persistent hover states, meaning the top-most link stays 'hovered' even when your cursor has moved down the list. */<br />
<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul a{ /* The persistent hover state does however create a global style for links even before they're hovered. Here we undo these effects. */<br />
background:#121212;<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul li a:hover{ /* Here we define the most explicit hover states--what happens when you hover each individual link. */<br />
background:#d4d4d4;<br />
text-decoration:none;<br />
}<br />
</style><br />
<!-- *** End of Menubar CSS style code from MIT, Thankyouuu :D *** --><br />
<br />
<div id="border-top"></div> <br />
<br />
<!-- *** Percobaan kode menu dropdown MIT *** --><br />
<ul class="nav"><br />
<br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia">HOME</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Team">TEAM</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Member">Members</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Lab">Our Lab</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Gallery">Gallery</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Project">PROJECT</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Description">Description</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Device">Device</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Parts">Parts</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Abstract">Abstract</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/JudgingForm">Judging Form</a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Notebook">NOTEBOOK</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/July">July</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/August">August</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/September">September</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices">HUMAN PRACTICES</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Sponsors">SPONSORS</a></li><br />
</ul><br />
<hr color:"white";> <br />
</html></div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/ProjectTeam:UI-Indonesia/Project2013-09-27T22:19:08Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br><br />
<br><br />
<h1>Page Underconstruction</h1> <br />
<!-- <h1><span id= "Projects">Project Description</span></h1><br />
<p><br />
<br />
Experimental<br><br />
<br>Tuberculosis (TB) is a worldwide major health problem. One third of the world’s population are estimated to be infected with Mycobacterium tuberculosis. It is the second major killer of adults after cardiovascular disease which kills 69.541 people and infect another 2.873.000 people anually. The absence of reliable diagnostic tool in suburban area, where TB cases are most likely found, is still a great obstacle in TB eradication effort.</br><br />
<br>Indonesia is one of the high burden countries for TB. The currently used method for detecting TB, the acid fast bacilli smear, has several difficulties such as getting the sputum (instead of saliva), and training the laboratory personnel to do the test. Based on this problem, UI iGEM team are trying to create a reliable, portable, and easy to use diagnostic tool for detecting TB. We are constructing a biosensor consist of highly specific antibody bound to a reporter to detect the presence of protein Ag85, a novel TB biomarker.</br><br />
<br>Here in this project, we are going to create a biosensor that will detect the presence of antigen 85, a potential tuberculosis biomarker. Antigen 85 is found in sputum, urine and most abundantly, blood serum of TB patient. Hence, blood serum of TB suspect will be used for the test. This biosensor designed to be able to detect TB more specifically than the existed method. The biosensor itself utilize split betagalactosidase combined with scFv (single chain fragment variable).</br><br />
<br>To complete the biosensor we created, characterization method to analyze the presence of Antigen 85 qualitatively and quantitatively already constructed. Characterization method is easy enough so this biosensor could be used as a good kit for TB detection in suburban area. </br> ---><br />
<br />
<br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/TeamTeam:UI-Indonesia/Team2013-09-27T22:17:03Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<br><br />
<br><br />
<br><br />
<h1><span id= "The_Great_Team"; style="color:white";>The Great Team</span></h1><br />
<br />
<img src="http://i1273.photobucket.com/albums/y401/motegum/MainPageWelcomeImage_zps8dba277c.jpg" border="0" alt=" photo MainPageWelcomeImage_zps8dba277c.jpg"/><br />
<div style="background-color:white";><br />
<h3><span id= "Lala">Muhammad Hanifi</span></h3><br />
<p><i>Profile currently unavailable</i></p><br />
<br />
<h3><span id= "Lili">Achmad Danny Gazali</span></h3><br />
<p><i>Profile currently unavailable</i></p><br />
<br />
<h3><span id= "Lulu">Dwiantari Satyapertiwi</span></h3><br />
<p><i>Profile currently unavailable</i></p><br />
<br />
<h3><span id= "Lulu">Mohamad Teguh Gumelar</span></h3><br />
<p><i>Profile currently unavailable</i></p><br />
<br />
<h3><span id= "Lulu">Putu Ayuwidia Ekaputri</span></h3><br />
<p><i>Profile currently unavailable</i></p><br />
<br />
<h3><span id= "Lulu">Rahdi Dewin Marzaini</span></h3><br />
<p><i>Profile currently unavailable</i></p><br />
<br />
<h3><span id= "Lulu">Taufik Hidayat Abdullah</span></h3><br />
<p><i>Profile currently unavailable</i></p><br />
<br />
<h3><span id= "Lulu">Wisnu Tafroji</span></h3><br />
<p><i>Profile currently unavailable</i></p><br />
<br />
</div><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/GalleryTeam:UI-Indonesia/Gallery2013-09-27T22:03:34Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/GalleryTeam:UI-Indonesia/Gallery2013-09-27T22:03:15Z<p>Taufik.ha: Created page with "<html> {{:Team:UI-Indonesia/header}} </html> {{:Team:UI-Indonesia/footer}}"</p>
<hr />
<div><html><br />
{{:Team:UI-Indonesia/header}}<br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/LabTeam:UI-Indonesia/Lab2013-09-27T22:02:39Z<p>Taufik.ha: Created page with "{{:Team:UI-Indonesia/header}} <html> </html> {{:Team:UI-Indonesia/footer}}"</p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/MemberTeam:UI-Indonesia/Member2013-09-27T22:02:05Z<p>Taufik.ha: Created page with "{{:Team:UI-Indonesia/header}} <html> </html> {{:Team:UI-Indonesia/footer}}"</p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/HumanPracticesTeam:UI-Indonesia/HumanPractices2013-09-27T21:55:20Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<body bgcolor="#efefef"><br />
<br />
<br><br><h1>Human Practice</h1></br><br />
<br>Our human practice mainly focused on the mission to socialize synthetic biology from some different perspective. We decided High School Students to be our main target since they are the young generation who will rule the future. Synthetic biology depends on the people who used it. Hence, describing some different perspective about it will give a big impact to their decision in defining synthetic biology.</br><br />
<br>Students targeted are students in High School around Jakarta. Their knowledge are about higher than other area, and synthetic biology facilities in those area are provided fairly. Hence, they have a big probability to deal with synthetic biology in the future. We have designed our human practice so that the students will get information directly in balance. We decided to use a simple learning kit we made by ourselves, to explain our project and also synthetic biology. The kit produced created simply so High School teacher will be able to use it in the class, for the next students in the next years.</br><br />
<br>In human practice, we also gather opinions and suggestions from the expert in synthetic biology to be discussed and illustrate synthetic biology in a whole. We will also use those opinions to fill the gap between student’s knowledge and complexity of our project. We also use artificial options to present the big impact of our project specifically, and synthetic biology generally. Not in order to make a good opinion about it, but to build a fair perspective about it.</br><br />
<br />
</body><br />
<br />
<br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/HumanPracticesTeam:UI-Indonesia/HumanPractices2013-09-27T21:52:49Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<style><br />
body<br />
{<br />
background-color:#efefef;<br />
}<br />
</style><br />
<body><br />
<br />
<br><br><h1>Human Practice</h1></br><br />
<br>Our human practice mainly focused on the mission to socialize synthetic biology from some different perspective. We decided High School Students to be our main target since they are the young generation who will rule the future. Synthetic biology depends on the people who used it. Hence, describing some different perspective about it will give a big impact to their decision in defining synthetic biology.</br><br />
<br>Students targeted are students in High School around Jakarta. Their knowledge are about higher than other area, and synthetic biology facilities in those area are provided fairly. Hence, they have a big probability to deal with synthetic biology in the future. We have designed our human practice so that the students will get information directly in balance. We decided to use a simple learning kit we made by ourselves, to explain our project and also synthetic biology. The kit produced created simply so High School teacher will be able to use it in the class, for the next students in the next years.</br><br />
<br>In human practice, we also gather opinions and suggestions from the expert in synthetic biology to be discussed and illustrate synthetic biology in a whole. We will also use those opinions to fill the gap between student’s knowledge and complexity of our project. We also use artificial options to present the big impact of our project specifically, and synthetic biology generally. Not in order to make a good opinion about it, but to build a fair perspective about it.</br><br />
<br />
</body><br />
<br />
<br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/HumanPracticesTeam:UI-Indonesia/HumanPractices2013-09-27T21:39:32Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<body background-color="white";><br />
<br><br><h1>Human Practice</h1></br><br />
<br>Our human practice mainly focused on the mission to socialize synthetic biology from some different perspective. We decided High School Students to be our main target since they are the young generation who will rule the future. Synthetic biology depends on the people who used it. Hence, describing some different perspective about it will give a big impact to their decision in defining synthetic biology.</br><br />
<br>Students targeted are students in High School around Jakarta. Their knowledge are about higher than other area, and synthetic biology facilities in those area are provided fairly. Hence, they have a big probability to deal with synthetic biology in the future. We have designed our human practice so that the students will get information directly in balance. We decided to use a simple learning kit we made by ourselves, to explain our project and also synthetic biology. The kit produced created simply so High School teacher will be able to use it in the class, for the next students in the next years.</br><br />
<br>In human practice, we also gather opinions and suggestions from the expert in synthetic biology to be discussed and illustrate synthetic biology in a whole. We will also use those opinions to fill the gap between student’s knowledge and complexity of our project. We also use artificial options to present the big impact of our project specifically, and synthetic biology generally. Not in order to make a good opinion about it, but to build a fair perspective about it.</br><br />
<br />
</body><br />
<br />
<br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/HumanPracticesTeam:UI-Indonesia/HumanPractices2013-09-27T21:37:40Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br><br><h1>Human Practice</h1></br><br />
<br>Our human practice mainly focused on the mission to socialize synthetic biology from some different perspective. We decided High School Students to be our main target since they are the young generation who will rule the future. Synthetic biology depends on the people who used it. Hence, describing some different perspective about it will give a big impact to their decision in defining synthetic biology.</br><br />
<br>Students targeted are students in High School around Jakarta. Their knowledge are about higher than other area, and synthetic biology facilities in those area are provided fairly. Hence, they have a big probability to deal with synthetic biology in the future. We have designed our human practice so that the students will get information directly in balance. We decided to use a simple learning kit we made by ourselves, to explain our project and also synthetic biology. The kit produced created simply so High School teacher will be able to use it in the class, for the next students in the next years.</br><br />
<br>In human practice, we also gather opinions and suggestions from the expert in synthetic biology to be discussed and illustrate synthetic biology in a whole. We will also use those opinions to fill the gap between student’s knowledge and complexity of our project. We also use artificial options to present the big impact of our project specifically, and synthetic biology generally. Not in order to make a good opinion about it, but to build a fair perspective about it.</br><br />
<br />
<br />
<br />
<br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/SponsorsTeam:UI-Indonesia/Sponsors2013-09-27T21:37:17Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<br><br />
<br><br />
<h1>Page Underconstruction</h1><br />
<br />
<br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-IndonesiaTeam:UI-Indonesia2013-09-27T21:36:51Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<html><br />
<br />
<div id= "section" style= "width: 960px; height: auto; text-align: justify;"><br />
<div style="text-align: center;"><br />
<br />
<br><br><br><br />
<br />
<h3><span style="font-family: Georgia,&quot;Times New Roman&quot;,serif; color:white;"> <i>Our goal is to create a new, easily used and sensitive method in detecting Tuberculosis</i> </span> </h3><br />
<br />
</div> <br />
</div><br />
<br />
<style><br />
td {padding:15px;align:center;}<br />
</style><br />
<br><br><br><br />
<table style="margin-left:120px; margin-right:120px; background-color:transparent"><br />
<tr><br />
<br />
<td><img src="http://i1273.photobucket.com/albums/y401/motegum/BlueIvyicontransparan_zps28755a8e.png" border="0" alt="Blue ivy icon - transparent photo BlueIvyicontransparan_zps28755a8e.png"/><br />
</td><br />
<br />
<td bgcolor="aqua"><br />
<h1>Project Description</h1><br />
<br />
<p>Tuberculosis (TB) is a worldwide major health problem. One third of the world’s population are estimated to be infected with Mycobacterium tuberculosis. It is the second major killer of adults after cardiovascular disease which kills 69.541 people and infect another 2.873.000 people anually. The absence of reliable diagnostic tool in suburban area, where TB cases are most likely found, is still a great obstacle in TB eradication effort.</p><br />
<br />
<p>Indonesia is one of the high burden countries for TB. The currently used method for detecting TB, the acid fast bacilli smear, has several difficulties such as getting the sputum (instead of saliva), and training the laboratory personnel to do the test. Based on this problem, UI iGEM team are trying to create a reliable, portable, and easy to use diagnostic tool for detecting TB. We are constructing a biosensor consist of highly specific antibody bound to a reporter to detect the presence of protein Ag85, a novel TB biomarker)</p><br />
<br />
<p>We are going to make a biosensor that will detect the presence of antigen 85, a potential tuberculosis biomarker. Antigen 85 is found in sputum, urine and most abundantly, blood serum of TB patient. Hence, blood serum of TB suspect will be used for the test. Positive result will be indicated with easy to detect blue color, when it’s negative, no response will be observed.</p><br />
</td><br />
<br />
</tr><br />
</table> <br />
<br />
<br />
</html><br />
{{:Team:UI-Indonesia/footer}}</div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/footerTeam:UI-Indonesia/footer2013-09-27T21:34:46Z<p>Taufik.ha: Created page with "<html> <!-- footer --> <div id= "footer" style="backgound-color: #b0c4de; width: 960px; height: auto; text-align:center"> <br> <br> <br> <br> <hr> <h3> <span style="fo..."</p>
<hr />
<div><html> <!-- footer --><br />
<div id= "footer" style="backgound-color: #b0c4de; width: 960px; height: auto; text-align:center"><br />
<br><br />
<br><br />
<br><br />
<br><br />
<hr><br />
<br />
<h3><br />
<span style="font-family: Georgia,&quot;Times New Roman&quot;,serif;color:white"><br />
<i>Supported by</i><br />
</span><br />
</h3><br />
<br />
<img src="http://i1273.photobucket.com/albums/y401/motegum/Supports_zps8f192e97.jpg" width="640" height="142" "border="0" alt=" photo Supports_zps8f192e97.jpg"><br />
<br />
</div><br />
</html></div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/headerTeam:UI-Indonesia/header2013-09-27T21:31:51Z<p>Taufik.ha: </p>
<hr />
<div><!-- *** Mencoba buat template isinya header, menu, background *** --><br />
<html><br />
<style><br />
body{position:absolute; top:0px; width:100%; height:100%;}<br />
<br />
#contentSub {<br />
display:none;<br />
}<br />
body {<br />
background: #fff url('https://static.igem.org/mediawiki/2013/c/c1/Bg3ui.jpg') no-repeat fixed;<br />
background-size: 1350px ;<br />
background-position: center top;<br />
}<br />
<br />
#top-section{<br />
height:0px;<br />
border:none;<br />
width:980px;<br />
margin:0 auto;<br />
padding:0 0 0 0;<br />
background-color:transparent;<br />
overflow:hide;<br />
<br />
}<br />
#p-logo{display:none;}<br />
#search-controls{display:none;}<br />
#top{display:none;}<br />
.firstHeading{display:none;}<br />
#footer-box{display:none;}<br />
#catlinks{display:none;}<br />
#content {background-color:transparent; border-left-width:0px; border-right-width:0px; padding: 0px 5px 0px 5px}<br />
</style><br />
<body><br />
<!-- header of the page --><br />
<header><br />
<div id= "header" style= "float: none; width: 960px;<br />
height: auto;"><br />
<br />
<!-- *** Mulai kode gambar header oleh Teguh *** --><br />
<img src="https://static.igem.org/mediawiki/2013/6/67/Header_New_Design.png" border="0"><br />
<br />
<br />
<!-- *** Akhir kode gambar header oleh Teguh *** --><br />
<br />
</header><br />
<div><br />
<br />
<!-- *** Menu Style CSS From MIT 2012, Thankyou..!! :D *** --><br />
<style><br />
.nav{<br />
float:left;<br />
width:100%;<br />
margin:0;<br />
list-style:none;<br />
position:relative;<br />
z-index:5;<br />
<br />
/*CHANGE THIS TO CENTER*/<br />
padding:0 185px; /*12px;*/<br />
}<br />
<br />
.nav li{<br />
float:left;<br />
position:relative;<br />
display:inline;<br />
line-height: 2em;<br />
}<br />
<br />
.nav a{<br />
float:left;<br />
height:2em;<br />
padding: 0 1.3em;<br />
color:white;<br />
/*Padding: bottom, right, top, left */<br />
/*padding:0.2em 1.3em 0.2em 1.3em;*/<br />
border-right:1px solid white;<br />
white-space:nowrap;<br />
}<br />
<br />
.nav a:hover{<br />
color:#00FFFF !important;<br />
}<br />
<br />
/*--- DROPDOWN ---*/<br />
.nav ul{<br />
background:#fff; <br />
list-style:none;<br />
position:absolute;<br />
left:-9999px; /* Hide off-screen when not needed (this is more accessible than display:none;) */<br />
margin-top: 25px;<br />
margin-left:0;<br />
margin-right:0;<br />
<br />
}<br />
<br />
<br />
.nav ul li{<br />
padding-top:1px;<br />
}<br />
<br />
.nav ul a{<br />
padding-top: 3px;<br />
padding-bottom:3px; /*Introducing a padding between the li and the a give the illusion spaced items */<br />
white-space:normal;<br />
text-decoration:none;<br />
height:auto;<br />
width: 13em;<br />
line-height: 1.5em;<br />
}<br />
<br />
.nav li:hover ul{ /* Display the dropdown on hover */<br />
left:0; /* Bring back on-screen when needed */<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover a{ /* These create persistent hover states, meaning the top-most link stays 'hovered' even when your cursor has moved down the list. */<br />
<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul a{ /* The persistent hover state does however create a global style for links even before they're hovered. Here we undo these effects. */<br />
background:#121212;<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul li a:hover{ /* Here we define the most explicit hover states--what happens when you hover each individual link. */<br />
background:#d4d4d4;<br />
text-decoration:none;<br />
}<br />
</style><br />
<!-- *** End of Menubar CSS style code from MIT, Thankyouuu :D *** --><br />
<br />
<div id="border-top"></div> <br />
<br />
<!-- *** Percobaan kode menu dropdown MIT *** --><br />
<ul class="nav"><br />
<br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia">HOME</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Team">TEAM</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Member">Members</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Lab">Our Lab</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Gallery">Gallery</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Project">PROJECT</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Description">Description</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Device">Device</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Parts">Parts</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Abstract">Abstract</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/JudgingForm">Judging Form</a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Notebook">NOTEBOOK</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices">HUMAN PRACTICES</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Sponsors">SPONSORS</a></li><br />
</ul><br />
<hr color:"white";> <br />
</html></div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/headerTeam:UI-Indonesia/header2013-09-27T19:07:59Z<p>Taufik.ha: </p>
<hr />
<div><!-- *** Mencoba buat template isinya header, menu, background *** --><br />
<html><br />
<style><br />
body{position:absolute; top:0px; width:100%; height:100%;}<br />
<br />
#contentSub {<br />
display:none;<br />
}<br />
body {<br />
background: #fff url('https://static.igem.org/mediawiki/2013/c/c1/Bg3ui.jpg') no-repeat fixed;<br />
background-size: 1350px ;<br />
background-position: center top;<br />
}<br />
<br />
#top-section{<br />
height:0px;<br />
border:none;<br />
width:980px;<br />
margin:0 auto;<br />
padding:0 0 0 0;<br />
background-color:transparent;<br />
overflow:hide;<br />
<br />
}<br />
#p-logo{display:none;}<br />
#search-controls{display:none;}<br />
#top{display:none;}<br />
.firstHeading{display:none;}<br />
#footer-box{display:none;}<br />
#catlinks{display:none;}<br />
#content {background-color:transparent; border-left-width:0px; border-right-width:0px; padding: 0px 5px 0px 5px}<br />
</style><br />
<body><br />
<!-- header of the page --><br />
<header><br />
<div id= "header" style= "float: none; width: 960px;<br />
height: auto;"><br />
<br />
<!-- *** Mulai kode gambar header oleh Teguh *** --><br />
<img src="https://static.igem.org/mediawiki/2013/6/67/Header_New_Design.png" border="0"><br />
<br />
<br />
<!-- *** Akhir kode gambar header oleh Teguh *** --><br />
<br />
</header><br />
<div><br />
<br />
<!-- *** Menu Style CSS From MIT 2012, Thankyou..!! :D *** --><br />
<style><br />
#menubar { <br />
background-color: white; <br />
}<br />
#menubar ul li a { <br />
color: #999999; }<br />
.right-menu li a {<br />
color: black;<br />
background-color: white;<br />
}<br />
<br />
.nav{<br />
float:left;<br />
width:100%;<br />
margin:0;<br />
list-style:none;<br />
position:relative;<br />
background-image:url(https://static.igem.org/mediawiki/2013/4/4a/Menubar_cokelat.png);background-repeat:no-repeat;<br />
z-index:5;<br />
<br />
/*CHANGE THIS TO CENTER*/<br />
padding:0 12px; /*12px;*/<br />
}<br />
<br />
.nav li{<br />
float:left;<br />
position:relative;<br />
display:inline;<br />
line-height: 2em;<br />
}<br />
<br />
.nav a{<br />
float:left;<br />
height:2em;<br />
padding: 0 1.3em;<br />
color:white;<br />
/*Padding: bottom, right, top, left */<br />
/*padding:0.2em 1.3em 0.2em 1.3em;*/<br />
border-right:1px solid white;<br />
white-space:nowrap;<br />
}<br />
<br />
.nav a:hover{<br />
color:#00FFFF !important;<br />
}<br />
<br />
/*--- DROPDOWN ---*/<br />
.nav ul{<br />
background:#fff; <br />
list-style:none;<br />
position:absolute;<br />
left:-9999px; /* Hide off-screen when not needed (this is more accessible than display:none;) */<br />
margin-top: 25px;<br />
margin-left:0;<br />
margin-right:0;<br />
<br />
}<br />
<br />
<br />
.nav ul li{<br />
padding-top:1px;<br />
}<br />
<br />
.nav ul a{<br />
padding-top: 3px;<br />
padding-bottom:3px; /*Introducing a padding between the li and the a give the illusion spaced items */<br />
white-space:normal;<br />
text-decoration:none;<br />
height:auto;<br />
width: 13em;<br />
line-height: 1.5em;<br />
}<br />
<br />
.nav li:hover ul{ /* Display the dropdown on hover */<br />
left:0; /* Bring back on-screen when needed */<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover a{ /* These create persistent hover states, meaning the top-most link stays 'hovered' even when your cursor has moved down the list. */<br />
background:#666666;<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul a{ /* The persistent hover state does however create a global style for links even before they're hovered. Here we undo these effects. */<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul li a:hover{ /* Here we define the most explicit hover states--what happens when you hover each individual link. */<br />
background:#993333;<br />
text-decoration:none;<br />
}<br />
</style><br />
<!-- *** End of Menubar CSS style code from MIT, Thankyouuu :D *** --><br />
<br />
<div id="border-top"></div> <br />
<br />
<!-- *** Percobaan kode menu dropdown MIT *** --><br />
<ul class="nav"><br />
<br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia">HOME</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Team">TEAM</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Member">Members</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Lab">Our Lab</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Gallery">Gallery</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Project">PROJECT</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Description">Description</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Device">Device</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Parts">Parts</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Abstract">Abstract</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/JudgingForm">Judging Form</a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Notebook">NOTEBOOK</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices">HUMAN PRACTICES</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Sponsors">SPONSORS</a></li><br />
<br />
</html></div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/headerTeam:UI-Indonesia/header2013-09-27T18:56:20Z<p>Taufik.ha: </p>
<hr />
<div><!-- *** Mencoba buat template isinya header, menu, background *** --><br />
<html><br />
<style><br />
body{position:absolute; top:0px; width:100%; height:100%;}<br />
<br />
#contentSub {<br />
display:none;<br />
}<br />
body {<br />
background: #fff url('https://static.igem.org/mediawiki/2013/c/c1/Bg3ui.jpg') no-repeat fixed;<br />
background-size: 1350px ;<br />
background-position: center top;<br />
}<br />
<br />
#top-section{<br />
height:0px;<br />
border:none;<br />
width:980px;<br />
margin:0 auto;<br />
padding:0 0 0 0;<br />
background-color:transparent;<br />
overflow:hide;<br />
<br />
}<br />
#p-logo{display:none;}<br />
#search-controls{display:none;}<br />
#top{display:none;}<br />
.firstHeading{display:none;}<br />
#footer-box{display:none;}<br />
#catlinks{display:none;}<br />
#content {background-color:transparent; border-left-width:0px; border-right-width:0px; padding: 0px 5px 0px 5px}<br />
</style><br />
<body><br />
<!-- header of the page --><br />
<header><br />
<div id= "header" style= "float: none; width: 960px;<br />
height: auto;"><br />
<br />
<!-- *** Mulai kode gambar header oleh Teguh *** --><br />
<img src="https://static.igem.org/mediawiki/2013/6/67/Header_New_Design.png" border="0"><br />
<br />
<br />
<!-- *** Akhir kode gambar header oleh Teguh *** --><br />
<br />
</header><br />
<div><br />
<br />
<!-- *** Menu Style CSS From MIT 2012, Thankyou..!! :D *** --><br />
<style><br />
#menubar { <br />
background-color: white; <br />
}<br />
#menubar ul li a { <br />
color: #999999; }<br />
.right-menu li a {<br />
color: black;<br />
background-color: white;<br />
}<br />
<br />
.nav{<br />
float:left;<br />
width:100%;<br />
margin:0;<br />
list-style:none;<br />
position:relative;<br />
z-index:5;<br />
<br />
/*CHANGE THIS TO CENTER*/<br />
padding:0 12px; /*12px;*/<br />
}<br />
<br />
.nav li{<br />
float:left;<br />
position:relative;<br />
display:inline;<br />
line-height: 2em;<br />
}<br />
<br />
.nav a{<br />
float:left;<br />
height:2em;<br />
padding: 0 1.3em;<br />
color:white;<br />
background-image:url(https://static.igem.org/mediawiki/2013/4/4a/Menubar_cokelat.png);<br />
/*Padding: bottom, right, top, left */<br />
/*padding:0.2em 1.3em 0.2em 1.3em;*/<br />
border-right:1px solid white;<br />
white-space:nowrap;<br />
}<br />
<br />
.nav a:hover{<br />
color:#00FFFF !important;<br />
}<br />
<br />
/*--- DROPDOWN ---*/<br />
.nav ul{<br />
background:#fff; <br />
list-style:none;<br />
position:absolute;<br />
left:-9999px; /* Hide off-screen when not needed (this is more accessible than display:none;) */<br />
margin-top: 25px;<br />
margin-left:0;<br />
margin-right:0;<br />
<br />
}<br />
<br />
<br />
.nav ul li{<br />
padding-top:1px;<br />
}<br />
<br />
.nav ul a{<br />
padding-top: 3px;<br />
padding-bottom:3px; /*Introducing a padding between the li and the a give the illusion spaced items */<br />
white-space:normal;<br />
text-decoration:none;<br />
height:auto;<br />
width: 13em;<br />
line-height: 1.5em;<br />
}<br />
<br />
.nav li:hover ul{ /* Display the dropdown on hover */<br />
left:0; /* Bring back on-screen when needed */<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover a{ /* These create persistent hover states, meaning the top-most link stays 'hovered' even when your cursor has moved down the list. */<br />
background:#666666;<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul a{ /* The persistent hover state does however create a global style for links even before they're hovered. Here we undo these effects. */<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul li a:hover{ /* Here we define the most explicit hover states--what happens when you hover each individual link. */<br />
background:#993333;<br />
text-decoration:none;<br />
}<br />
</style><br />
<!-- *** End of Menubar CSS style code from MIT, Thankyouuu :D *** --><br />
<br />
<div id="border-top"></div> <br />
<br />
<!-- *** Percobaan kode menu dropdown MIT *** --><br />
<ul class="nav"><br />
<br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia">HOME</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Team">TEAM</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Member">Members</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Lab">Our Lab</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Gallery">Gallery</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Project">PROJECT</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Description">Description</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Device">Device</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Parts">Parts</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Abstract">Abstract</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/JudgingForm">Judging Form</a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Notebook">NOTEBOOK</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices">HUMAN PRACTICES</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Sponsors">SPONSORS</a></li><br />
<br />
</html></div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/headerTeam:UI-Indonesia/header2013-09-27T18:39:24Z<p>Taufik.ha: </p>
<hr />
<div><!-- *** Mencoba buat template isinya header, menu, background *** --><br />
<html><br />
<style><br />
body{position:absolute; top:0px; width:100%; height:100%;}<br />
<br />
#contentSub {<br />
display:none;<br />
}<br />
body {<br />
background: #fff url('https://static.igem.org/mediawiki/2013/c/c1/Bg3ui.jpg') no-repeat fixed;<br />
background-size: 1350px ;<br />
background-position: center top;<br />
}<br />
<br />
#top-section{<br />
height:0px;<br />
border:none;<br />
width:980px;<br />
margin:0 auto;<br />
padding:0 0 0 0;<br />
background-color:transparent;<br />
overflow:hide;<br />
<br />
}<br />
#p-logo{display:none;}<br />
#search-controls{display:none;}<br />
#top{display:none;}<br />
.firstHeading{display:none;}<br />
#footer-box{display:none;}<br />
#catlinks{display:none;}<br />
#content {background-color:transparent; border-left-width:0px; border-right-width:0px; padding: 0px 5px 0px 5px}<br />
</style><br />
<body><br />
<!-- header of the page --><br />
<header><br />
<div id= "header" style= "float: none; width: 960px;<br />
height: auto;"><br />
<br />
<!-- *** Mulai kode gambar header oleh Teguh *** --><br />
<img src="https://static.igem.org/mediawiki/2013/6/67/Header_New_Design.png" border="0"><br />
<br />
<br />
<!-- *** Akhir kode gambar header oleh Teguh *** --><br />
<br />
</header><br />
<div><br />
<br />
<!-- *** Menu Style CSS From MIT 2012, Thankyou..!! :D *** --><br />
<style><br />
#menubar { <br />
background-color: white; <br />
}<br />
#menubar ul li a { <br />
color: #999999; }<br />
.right-menu li a {<br />
color: black;<br />
background-color: white;<br />
}<br />
<br />
.nav{<br />
float:left;<br />
width:100%;<br />
margin:0;<br />
list-style:none;<br />
position:relative;<br />
z-index:5;<br />
/*CHANGE THIS TO CENTER*/<br />
padding:0 12px; /*12px;*/<br />
}<br />
<br />
.nav li{<br />
float:left;<br />
position:relative;<br />
display:inline;<br />
line-height: 2em;<br />
}<br />
<br />
.nav a{<br />
float:left;<br />
height:2em;<br />
padding: 0 1.3em;<br />
color:white;<br />
background-color:#666666;<br />
/*Padding: bottom, right, top, left */<br />
/*padding:0.2em 1.3em 0.2em 1.3em;*/<br />
border-right:1px solid white;<br />
white-space:nowrap;<br />
}<br />
<br />
.nav a:hover{<br />
background-color:#993333 !important;<br />
}<br />
<br />
/*--- DROPDOWN ---*/<br />
.nav ul{<br />
background:#fff; /* Adding a background makes the dropdown work properly in IE7+. Make this as close to your page's background as possible (i.e. white page == white background). */<br />
background:rgba(255,255,255,0); /* But! Let's make the background fully transparent where we can, we don't actually want to see it if we can help it... */<br />
list-style:none;<br />
position:absolute;<br />
left:-9999px; /* Hide off-screen when not needed (this is more accessible than display:none;) */<br />
margin-top: 25px;<br />
margin-left:0;<br />
margin-right:0;<br />
<br />
}<br />
<br />
<br />
.nav ul li{<br />
padding-top:1px;<br />
}<br />
<br />
.nav ul a{<br />
padding-top: 3px;<br />
padding-bottom:3px; /*Introducing a padding between the li and the a give the illusion spaced items */<br />
white-space:normal;<br />
text-decoration:none;<br />
height:auto;<br />
width: 13em;<br />
line-height: 1.5em;<br />
}<br />
<br />
.nav li:hover ul{ /* Display the dropdown on hover */<br />
left:0; /* Bring back on-screen when needed */<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover a{ /* These create persistent hover states, meaning the top-most link stays 'hovered' even when your cursor has moved down the list. */<br />
background:#666666;<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul a{ /* The persistent hover state does however create a global style for links even before they're hovered. Here we undo these effects. */<br />
text-decoration:none;<br />
}<br />
<br />
.nav li:hover ul li a:hover{ /* Here we define the most explicit hover states--what happens when you hover each individual link. */<br />
background:#993333;<br />
text-decoration:none;<br />
}<br />
</style><br />
<!-- *** End of Menubar CSS style code from MIT, Thankyouuu :D *** --><br />
<br />
<div id="border-top"></div> <br />
<br />
<!-- *** Percobaan kode menu dropdown MIT *** --><br />
<ul class="nav"><br />
<br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia">HOME</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Team">TEAM</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Member">Members</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Lab">Our Lab</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Gallery">Gallery</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Project">PROJECT</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Description">Description</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Device">Device</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Parts">Parts</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Abstract">Abstract</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/JudgingForm">Judging Form</a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Notebook">NOTEBOOK</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices">HUMAN PRACTICES</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Sponsors">SPONSORS</a></li><br />
<br />
</html></div>Taufik.hahttp://2013.igem.org/Team:UI-IndonesiaTeam:UI-Indonesia2013-09-27T03:51:13Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<br />
<html><br />
<br />
<div id= "section" style= "width: 960px; height: auto; text-align: justify;"><br />
<div style="text-align: center;"><br />
<br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br />
<h3><span style="font-family: Georgia,&quot;Times New Roman&quot;,serif; color:white;"> <i>Our goal is to create a new, easily used and sensitive method in detecting Tuberculosis</i> </span> </h3><br />
<br />
</div> <br />
</div><br />
<br />
<style><br />
td {padding:15px;align:center;}<br />
</style><br />
<br />
<table style="margin-left:120px; margin-right:120px; background-color:transparent"><br />
<tr><br />
<br />
<td><img src="http://i1273.photobucket.com/albums/y401/motegum/BlueIvyicontransparan_zps28755a8e.png" border="0" alt="Blue ivy icon - transparent photo BlueIvyicontransparan_zps28755a8e.png"/><br />
</td><br />
<br />
<td bgcolor="aqua"><br />
<h1>Project Description</h1><br />
<br />
<p>Tuberculosis (TB) is a worldwide major health problem. One third of the world’s population are estimated to be infected with Mycobacterium tuberculosis. It is the second major killer of adults after cardiovascular disease which kills 69.541 people and infect another 2.873.000 people anually. The absence of reliable diagnostic tool in suburban area, where TB cases are most likely found, is still a great obstacle in TB eradication effort.</p><br />
<br />
<p>Indonesia is one of the high burden countries for TB. The currently used method for detecting TB, the acid fast bacilli smear, has several difficulties such as getting the sputum (instead of saliva), and training the laboratory personnel to do the test. Based on this problem, UI iGEM team are trying to create a reliable, portable, and easy to use diagnostic tool for detecting TB. We are constructing a biosensor consist of highly specific antibody bound to a reporter to detect the presence of protein Ag85, a novel TB biomarker)</p><br />
<br />
<p>We are going to make a biosensor that will detect the presence of antigen 85, a potential tuberculosis biomarker. Antigen 85 is found in sputum, urine and most abundantly, blood serum of TB patient. Hence, blood serum of TB suspect will be used for the test. Positive result will be indicated with easy to detect blue color, when it’s negative, no response will be observed.</p><br />
</td><br />
<br />
</tr><br />
</table> <br />
<br />
<!-- footer --><br />
<div id= "footer" style="backgound-color: #b0c4de; width: 960px; height: auto; text-align:center"><br />
<br><br />
<br><br />
<br><br />
<br><br />
<hr><br />
<br />
<h3><br />
<span style="font-family: Georgia,&quot;Times New Roman&quot;,serif;color:white"><br />
<i>Supported by</i><br />
</span><br />
</h3><br />
<br />
<img src="http://i1273.photobucket.com/albums/y401/motegum/Supports_zps8f192e97.jpg" width="640" height="142" "border="0" alt=" photo Supports_zps8f192e97.jpg"><br />
<br />
</div><br />
<br />
</html></div>Taufik.hahttp://2013.igem.org/Team:UI-IndonesiaTeam:UI-Indonesia2013-09-27T03:37:31Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<br />
<html><br />
<br />
<div id= "section" style= "width: 960px; height: auto; text-align: justify;"><br />
<div style="text-align: center;"><br />
<br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<h3> <span style="font-family: Georgia,&quot;Times New Roman&quot;,serif; color:white;"> <i>Our goal is to create a new, easily used and sensitive method in detecting Tuberculosis</i> </span> </h3><br />
<br />
</div> <br />
</div><br />
<style><br />
td {padding:15px;align:center;}<br />
</style><br />
<br />
<table style="margin-left:120px; margin-right:120px; background-color:transparent"><br />
<tr><br />
<br />
<td><img src="http://i1273.photobucket.com/albums/y401/motegum/BlueIvyicontransparan_zps28755a8e.png" border="0" alt="Blue ivy icon - transparent photo BlueIvyicontransparan_zps28755a8e.png"/></td><br />
<br />
<td bgcolor="aqua"><h1>Project Description</h1><br />
<p>Tuberculosis (TB) is a worldwide major health problem. One third of the world’s population are estimated to be infected with Mycobacterium tuberculosis. It is the second major killer of adults after cardiovascular disease which kills 69.541 people and infect another 2.873.000 people anually. The absence of reliable diagnostic tool in suburban area, where TB cases are most likely found, is still a great obstacle in TB eradication effort.</p><br />
<br />
<p>Indonesia is one of the high burden countries for TB. The currently used method for detecting TB, the acid fast bacilli smear, has several difficulties such as getting the sputum (instead of saliva), and training the laboratory personnel to do the test. Based on this problem, UI iGEM team are trying to create a reliable, portable, and easy to use diagnostic tool for detecting TB. We are constructing a biosensor consist of highly specific antibody bound to a reporter to detect the presence of protein Ag85, a novel TB biomarker)</p><br />
<br />
<p>We are going to make a biosensor that will detect the presence of antigen 85, a potential tuberculosis biomarker. Antigen 85 is found in sputum, urine and most abundantly, blood serum of TB patient. Hence, blood serum of TB suspect will be used for the test. Positive result will be indicated with easy to detect blue color, when it’s negative, no response will be observed.</p></td><br />
<br />
</tr><br />
<br />
</table> <br />
<br />
<!-- footer --><br />
<div id= "footer" style="backgound-color: #b0c4de; width: 960px; height: auto; text-align:center"><br />
<!-- footer-nya ini bisa diganti isinya --><br />
<br />
<br><br />
<br><br />
<br><br />
<br><br />
<hr><br />
<h3><br />
<span style="font-family: Georgia,&quot;Times New Roman&quot;,serif;color:white"><br />
<i>Supported by</i><br />
</span><br />
</h3><br />
<br />
<img src="http://i1273.photobucket.com/albums/y401/motegum/Supports_zps8f192e97.jpg" width="640" height="142" "border="0" alt=" photo Supports_zps8f192e97.jpg"><br />
</div><br />
</html></div>Taufik.hahttp://2013.igem.org/Team:UI-IndonesiaTeam:UI-Indonesia2013-09-27T03:31:39Z<p>Taufik.ha: </p>
<hr />
<div>{{:Team:UI-Indonesia/header}}<br />
<br />
<html><br />
<br />
<div id= "section" style= "width: 960px; height: auto; text-align: justify;"><br />
<div style="text-align: center;"><br />
<br />
<br><br />
<br><br />
<h2> <span style="font-family: Georgia,&quot;Times New Roman&quot;,serif; color:white;"> <i>Our goal is to create a new, easily used and sensitive method in detecting Tuberculosis</i></span> </h2><br />
<br />
</div> <br />
</div><br />
<br />
<br><br />
<br><br />
<br />
<style><br />
td {padding:15px;align:center;}<br />
</style><br />
<br />
<table style="margin-left:120px; margin-right:120px; background-color:transparent"><br />
<tr><br />
<br />
<td><img src="http://i1273.photobucket.com/albums/y401/motegum/BlueIvyicontransparan_zps28755a8e.png" border="0" alt="Blue ivy icon - transparent photo BlueIvyicontransparan_zps28755a8e.png"/></td><br />
<br />
<td bgcolor="aqua"><h1>Project Description</h1><br />
<p>Tuberculosis (TB) is a worldwide major health problem. One third of the world’s population are estimated to be infected with Mycobacterium tuberculosis. It is the second major killer of adults after cardiovascular disease which kills 69.541 people and infect another 2.873.000 people anually. The absence of reliable diagnostic tool in suburban area, where TB cases are most likely found, is still a great obstacle in TB eradication effort.</p><br />
<br />
<p>Indonesia is one of the high burden countries for TB. The currently used method for detecting TB, the acid fast bacilli smear, has several difficulties such as getting the sputum (instead of saliva), and training the laboratory personnel to do the test. Based on this problem, UI iGEM team are trying to create a reliable, portable, and easy to use diagnostic tool for detecting TB. We are constructing a biosensor consist of highly specific antibody bound to a reporter to detect the presence of protein Ag85, a novel TB biomarker)</p><br />
<br />
<p>We are going to make a biosensor that will detect the presence of antigen 85, a potential tuberculosis biomarker. Antigen 85 is found in sputum, urine and most abundantly, blood serum of TB patient. Hence, blood serum of TB suspect will be used for the test. Positive result will be indicated with easy to detect blue color, when it’s negative, no response will be observed.</p></td><br />
<br />
</tr><br />
<br />
</table> <br />
<br />
<!-- footer --><br />
<div id= "footer" style="backgound-color: #b0c4de; width: 960px; height: auto; text-align:center"><br />
<!-- footer-nya ini bisa diganti isinya --><br />
<br />
<br><br />
<br><br />
<br />
<h1><br />
<span style="font-family: Georgia,&quot;Times New Roman&quot;,serif;color:white"><br />
<i>Supported by</i><br />
</span><br />
</h1><br />
<br />
<img src="http://i1273.photobucket.com/albums/y401/motegum/Supports_zps8f192e97.jpg" width="640" height="142" "border="0" alt=" photo Supports_zps8f192e97.jpg"><br />
</div><br />
</html></div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/headerTeam:UI-Indonesia/header2013-09-27T03:27:42Z<p>Taufik.ha: </p>
<hr />
<div><!-- *** Mencoba buat template isinya header, menu, background *** --><br />
<html><br />
<style><br />
body{position:absolute; top:0px; width:100%; height:100%;}<br />
<br />
#contentSub {<br />
display:none;<br />
}<br />
body {<br />
background: #fff url('https://static.igem.org/mediawiki/2013/c/c1/Bg3ui.jpg') no-repeat fixed;<br />
background-size: 1350px ;<br />
background-position: center top;<br />
}<br />
<br />
#top-section{<br />
height:0px;<br />
border:none;<br />
width:980px;<br />
margin:0 auto;<br />
padding:0 0 0 0;<br />
background-color:transparent;<br />
overflow:hide;<br />
<br />
}<br />
#p-logo{display:none;}<br />
#search-controls{display:none;}<br />
#top{display:none;}<br />
.firstHeading{display:none;}<br />
#footer-box{display:none;}<br />
#catlinks{display:none;}<br />
#content {background-color:transparent; border-left-width:0px; border-right-width:0px; padding: 0px 5px 0px 5px}<br />
</style><br />
<body><br />
<!-- header of the page --><br />
<header><br />
<div id= "header" style= "float: none; width: 960px;<br />
height: auto;"><br />
<br />
<!-- *** Mulai kode gambar header oleh Teguh *** --><br />
<img src="http://i1273.photobucket.com/albums/y401/motegum/HeaderBarucompressed_zps7cc04662.jpg" border="0" alt="Header Baru Document photo HeaderBarucompressed_zps7cc04662.jpg"/><br />
<br />
<br />
<!-- *** Akhir kode gambar header oleh Teguh *** --><br />
<br />
</header><br />
<br />
<div><br />
<menu><br />
<style><br />
#nav { font-family: "Trebuchet MS", sans-serif; font-size: 15px; color: white;} <br />
#nav { background-color: #096f02; width: 845px; list-style: none; position:center; }<br />
#nav { float: left;}<br />
#nav li { float: left; margin: 0px; position:relative; }<br />
#nav a { display: block; padding: 5px 15px; text-decoration: none; color: white; border-style: none; }<br />
#nav a:hover { color: #f9fd92; }<br />
</style><br />
<br />
<div id="border-top"></div> <br />
<ul id="nav"><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia">HOME</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Team">TEAM</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Project">PROJECT</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices">HUMAN PRACTICES</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Notebook">NOTEBOOK</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Outreach">OUTREACH</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/About">ABOUT US</a></li><br />
</menu><br />
</div><br />
<br />
<br />
<br />
<!-- *** Percobaan ikon menu oleh Teguh *** --><br />
<!-- *** <div><br />
<a href="https://2013.igem.org/Team:UI-Indonesia"><img src="http://i1273.photobucket.com/albums/y401/motegum/Home_zps0c8ea51e.jpg" width="auto" height="70" border="0" alt=" photo ButtonHome_zps97ec748e.png"/>Home</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/Parts"><img src="http://i1273.photobucket.com/albums/y401/motegum/puzzle-vector_zps2833a421.jpg" width="auto" height="70" border="0" alt=" photo ButtonParts_zpscad1bf3f.png"/>Parts</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/Team"><img src="http://i1273.photobucket.com/albums/y401/motegum/Team_zpsc7153c46.jpg" width="auto" height="70" border="0" alt=" photo ButtonBackground2_zps5baf8876.png"/>Team</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/Project"><img src="http://i1273.photobucket.com/albums/y401/motegum/project_zps19a8e841.jpg" width="auto" height="70" border="0" alt=" photo ButtonBackground2_zps5baf8876.png"/>Project</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices"><img src="http://i1273.photobucket.com/albums/y401/motegum/HumanPractise_zpse76245c0.jpg" width="auto" height="70" border="0" alt=" photo ButtonBackground2_zps5baf8876.png"/>Human Practices</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/Notebook"><img src="http://i1273.photobucket.com/albums/y401/motegum/Notebook_zps2f87faf7.jpg" width="auto" height="70" border="0" alt=" photo ButtonBackground2_zps5baf8876.png"/>Notebook</a> *** --><br />
<!-- *** Percobaan ikon menu oleh Teguh *** --><br />
</div><br />
<br />
<br />
<br />
<br />
<!-- body navigation bar --><br />
<!-- di sini kodenya ada yang diambil dari wiki Edinburgh, yang dari contoh --><br />
<br />
<br />
<br />
<!-- body section --><br />
<div id= "section" style= "width: 960px; height: auto; text-align: justify"><br />
</html></div>Taufik.hahttp://2013.igem.org/File:Bg3ui.jpgFile:Bg3ui.jpg2013-09-27T03:27:02Z<p>Taufik.ha: </p>
<hr />
<div></div>Taufik.hahttp://2013.igem.org/Team:UI-Indonesia/headerTeam:UI-Indonesia/header2013-09-27T03:26:00Z<p>Taufik.ha: </p>
<hr />
<div><!-- *** Mencoba buat template isinya header, menu, background *** --><br />
<html><br />
<style><br />
body{position:absolute; top:0px; width:100%; height:100%;}<br />
<br />
#contentSub {<br />
display:none;<br />
}<br />
body {<br />
background: #fff url('https://static.igem.org/mediawiki/2013/2/20/Bg2ui.jpg') no-repeat fixed;<br />
background-size: 1350px ;<br />
background-position: center top;<br />
}<br />
<br />
#top-section{<br />
height:0px;<br />
border:none;<br />
width:980px;<br />
margin:0 auto;<br />
padding:0 0 0 0;<br />
background-color:transparent;<br />
overflow:hide;<br />
<br />
}<br />
#p-logo{display:none;}<br />
#search-controls{display:none;}<br />
#top{display:none;}<br />
.firstHeading{display:none;}<br />
#footer-box{display:none;}<br />
#catlinks{display:none;}<br />
#content {background-color:transparent; border-left-width:0px; border-right-width:0px; padding: 0px 5px 0px 5px}<br />
</style><br />
<body><br />
<!-- header of the page --><br />
<header><br />
<div id= "header" style= "float: none; width: 960px;<br />
height: auto;"><br />
<br />
<!-- *** Mulai kode gambar header oleh Teguh *** --><br />
<img src="http://i1273.photobucket.com/albums/y401/motegum/HeaderBarucompressed_zps7cc04662.jpg" border="0" alt="Header Baru Document photo HeaderBarucompressed_zps7cc04662.jpg"/><br />
<br />
<br />
<!-- *** Akhir kode gambar header oleh Teguh *** --><br />
<br />
</header><br />
<br />
<div><br />
<menu><br />
<style><br />
#nav { font-family: "Trebuchet MS", sans-serif; font-size: 15px; color: white;} <br />
#nav { background-color: #096f02; width: 845px; list-style: none; position:center; }<br />
#nav { float: left;}<br />
#nav li { float: left; margin: 0px; position:relative; }<br />
#nav a { display: block; padding: 5px 15px; text-decoration: none; color: white; border-style: none; }<br />
#nav a:hover { color: #f9fd92; }<br />
</style><br />
<br />
<div id="border-top"></div> <br />
<ul id="nav"><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia">HOME</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Team">TEAM</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Project">PROJECT</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices">HUMAN PRACTICES</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Notebook">NOTEBOOK</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/Outreach">OUTREACH</a></li><br />
<li><a href="https://2013.igem.org/Team:UI-Indonesia/About">ABOUT US</a></li><br />
</menu><br />
</div><br />
<br />
<br />
<br />
<!-- *** Percobaan ikon menu oleh Teguh *** --><br />
<!-- *** <div><br />
<a href="https://2013.igem.org/Team:UI-Indonesia"><img src="http://i1273.photobucket.com/albums/y401/motegum/Home_zps0c8ea51e.jpg" width="auto" height="70" border="0" alt=" photo ButtonHome_zps97ec748e.png"/>Home</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/Parts"><img src="http://i1273.photobucket.com/albums/y401/motegum/puzzle-vector_zps2833a421.jpg" width="auto" height="70" border="0" alt=" photo ButtonParts_zpscad1bf3f.png"/>Parts</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/Team"><img src="http://i1273.photobucket.com/albums/y401/motegum/Team_zpsc7153c46.jpg" width="auto" height="70" border="0" alt=" photo ButtonBackground2_zps5baf8876.png"/>Team</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/Project"><img src="http://i1273.photobucket.com/albums/y401/motegum/project_zps19a8e841.jpg" width="auto" height="70" border="0" alt=" photo ButtonBackground2_zps5baf8876.png"/>Project</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/HumanPractices"><img src="http://i1273.photobucket.com/albums/y401/motegum/HumanPractise_zpse76245c0.jpg" width="auto" height="70" border="0" alt=" photo ButtonBackground2_zps5baf8876.png"/>Human Practices</a><br />
<br />
<a href="https://2013.igem.org/Team:UI-Indonesia/Notebook"><img src="http://i1273.photobucket.com/albums/y401/motegum/Notebook_zps2f87faf7.jpg" width="auto" height="70" border="0" alt=" photo ButtonBackground2_zps5baf8876.png"/>Notebook</a> *** --><br />
<!-- *** Percobaan ikon menu oleh Teguh *** --><br />
</div><br />
<br />
<br />
<br />
<br />
<!-- body navigation bar --><br />
<!-- di sini kodenya ada yang diambil dari wiki Edinburgh, yang dari contoh --><br />
<br />
<br />
<br />
<!-- body section --><br />
<div id= "section" style= "width: 960px; height: auto; text-align: justify"><br />
</html></div>Taufik.hahttp://2013.igem.org/File:Bg2ui.jpgFile:Bg2ui.jpg2013-09-27T03:25:15Z<p>Taufik.ha: </p>
<hr />
<div></div>Taufik.ha