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Team:Hong Kong HKUST/notebook/mod4
From 2013.igem.org
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<div> | <div> | ||
<a href='#' class='head'>Week 1</a> | <a href='#' class='head'>Week 1</a> | ||
| - | <div class='content' align="left"> | + | <div class='content' align="left"> |
| + | <p> </p> | ||
| + | <p>Transformed iDUET101<br /> | ||
| + | Mini prepped transformant and do digestion check<br /> | ||
| + | Make glycerol stock out of iDUET101<br /> | ||
| + | Extract out Psb1C3 from BBa_J04450 using gel extraction with XbaI and PstI<br /> | ||
| + | Extract <i>E. coli</i> BW25113 genome</p> | ||
| + | <p> </p> | ||
</div> | </div> | ||
</div> | </div> | ||
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<div> | <div> | ||
<a href='#' class='head'>Week 2</a> | <a href='#' class='head'>Week 2</a> | ||
| - | <div class='content' align="left"> | + | <div class='content' align="left"> |
| + | <p> </p> | ||
| + | <p>Ligation of aceA and aceB with pSB1c3<br /> | ||
| + | Transformed aceA-pSB1C3 and aceB-pSB1C3<br /> | ||
| + | Screen out colonies for aceA-pSB1C3 and aceB-pSB1C3 and do digestion check<br /> | ||
| + | aceB-pSB1C3 confirmed and proceed with sequencing<br /> | ||
| + | aceB-pSB1C3 partial sequence confirmed</p> | ||
| + | <p> </p> | ||
</div> | </div> | ||
</div> | </div> | ||
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<div> | <div> | ||
<a href='#' class='head'>Week 3</a> | <a href='#' class='head'>Week 3</a> | ||
| - | <div class='content' align="left"> | + | <div class='content' align="left"> |
| + | <p> </p> | ||
| + | <p>Extracted out more Psb1c3 from BBa_J04450<br /> | ||
| + | PCR out aceA with standard prefix and suffix<br /> | ||
| + | Digestion of aceA with XbaI and PstI<br /> | ||
| + | Ligate with pSB1C3<br /> | ||
| + | Screen colonies and do digestion check<br /> | ||
| + | aceA-pSB1C3 confirmed and proceed with sequencing<br /> | ||
| + | aceA-pSB1C3 partial sequence confirmed<br /> | ||
| + | Help Chloe do digestion check for EF1a-pSB1C3</p> | ||
| + | <p> </p> | ||
</div> | </div> | ||
</div> | </div> | ||
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<div> | <div> | ||
<a href='#' class='head'>Week 4</a> | <a href='#' class='head'>Week 4</a> | ||
| - | <div class='content' align="left"> | + | <div class='content' align="left"> |
| + | <p> </p> | ||
| + | <p>Extraction of EF1a for Biobrick<br /> | ||
| + | Extract out more pSB1C3 from BBa_J04450<br /> | ||
| + | Digestion of EF1a with EcoRI and PstI<br /> | ||
| + | Ligation of EF1a with pSB1c3<br /> | ||
| + | Transformed EF1a-pSB1C3 construct<br /> | ||
| + | Screen colonies and do digestion check<br /> | ||
| + | Weird bands show up, do colony PCR to confirm whether EF1a is ligated<br /> | ||
| + | Colony PCR showed that EF1a is there<br /> | ||
| + | Colony PCR with VF2 and VR primers didn’t give any result</p> | ||
| + | <p> </p> | ||
</div> | </div> | ||
</div> | </div> | ||
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<div id="satu" align="center"> <h1>August 2013</h1> | <div id="satu" align="center"> <h1>August 2013</h1> | ||
Revision as of 17:12, 19 September 2013
June 2013
Design primers for aceA, aceB, MLS and EF1a extraction
Mini prepped pCMV/myc/mito
Extracted BW25113 genome
PCR out aceA and aceB for Biobrick
PCR out MLS for fuse with aceA and aceB
PCR out aceA and aceB to fuse with MLS
July 2013
Transformed iDUET101
Mini prepped transformant and do digestion check
Make glycerol stock out of iDUET101
Extract out Psb1C3 from BBa_J04450 using gel extraction with XbaI and PstI
Extract E. coli BW25113 genome
Ligation of aceA and aceB with pSB1c3
Transformed aceA-pSB1C3 and aceB-pSB1C3
Screen out colonies for aceA-pSB1C3 and aceB-pSB1C3 and do digestion check
aceB-pSB1C3 confirmed and proceed with sequencing
aceB-pSB1C3 partial sequence confirmed
Extracted out more Psb1c3 from BBa_J04450
PCR out aceA with standard prefix and suffix
Digestion of aceA with XbaI and PstI
Ligate with pSB1C3
Screen colonies and do digestion check
aceA-pSB1C3 confirmed and proceed with sequencing
aceA-pSB1C3 partial sequence confirmed
Help Chloe do digestion check for EF1a-pSB1C3
Extraction of EF1a for Biobrick
Extract out more pSB1C3 from BBa_J04450
Digestion of EF1a with EcoRI and PstI
Ligation of EF1a with pSB1c3
Transformed EF1a-pSB1C3 construct
Screen colonies and do digestion check
Weird bands show up, do colony PCR to confirm whether EF1a is ligated
Colony PCR showed that EF1a is there
Colony PCR with VF2 and VR primers didn’t give any result
