Team:Caltech/week3

From 2014.igem.org

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To figure out what went wrong during assembly of the combinatorial promoter constructs used in last week's TXTL workshop, we attempted to take out the LacI and TetR genes present in plasmids pAA001 and pAA002.
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To figure out what went wrong during assembly of the combinatorial promoter constructs used in last week's TXTL workshop, we attempted to take out the LacI and TetR genes present in plasmids pAA001.
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<ul><li>PCR extraction of linear fragments of pAA001 and pAA002 minus the lacI and tetR genes.</li>
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<ul><li>PCR extraction of linear fragments of pAA001 minus the lacI and tetR genes.</li>
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     <li>Recircularization of the truncated "backbone"+"combinatorial promoter construct" round-the-horn PCR</li>
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     <li>Recircularization of the truncated "backbone"+"combinatorial promoter construct" round-the-horn ligation</li>
     <li>Transformation of recircularized plasmids into JM109 competent cells</li>
     <li>Transformation of recircularized plasmids into JM109 competent cells</li>
     <li>Cells plated onto carbenicillin plates and incubated at 37&deg;C overnight</li>
     <li>Cells plated onto carbenicillin plates and incubated at 37&deg;C overnight</li>
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<ul><li>Colony PCR of colonies that grew up last night</li>
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     <li></li>
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     <li>Due to the long weekend, we decided that we will wait until next week to begin minipreps</li>
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Revision as of 22:51, 3 July 2014


Home Team Official Team Profile Project Parts Modeling Notebook Safety Attributions

Notebook

Overview

Week 1

Week 2

Week 3

Week 4

Week 5

Week 6

Week 7

Week 8

Week 9

Week 10

Week 11

Week 12

Week Three

Monday, 6/30/14
  • Discussion of fallout from Friday's meeting
  • More paper-reading to determine what's wrong with our new quorum-sensing system candidates
  • Designed geneblocks for our new systems and subgroup projects

Tuesday, 7/1/14
  • Lab cleanup. Reorganized -20 freezer samples.
  • World Cup watching. Tim Howard's a boss
  • Work on the team wiki while we wait for our next batch of geneblocks to ship

Wednesday, 7/2/14

 To figure out what went wrong during assembly of the combinatorial promoter constructs used in last week's TXTL workshop, we attempted to take out the LacI and TetR genes present in plasmids pAA001.
  • PCR extraction of linear fragments of pAA001 minus the lacI and tetR genes.
  • Recircularization of the truncated "backbone"+"combinatorial promoter construct" round-the-horn ligation
  • Transformation of recircularized plasmids into JM109 competent cells
  • Cells plated onto carbenicillin plates and incubated at 37°C overnight

In other news, we continued to work on the wiki and make it worse and worse......

Thursday, 7/3/14
  • Colony PCR of colonies that grew up last night
  • Due to the long weekend, we decided that we will wait until next week to begin minipreps

Friday, 7/4/14

 It's the 4th of July!!!
'Murica...or something like that, I guess...

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