Team:Caltech/week2

From 2014.igem.org

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<ul><li>Discussed alternatives to current ComQXPA system, including 2-cell system (not all in 1 cell)</li>
<ul><li>Discussed alternatives to current ComQXPA system, including 2-cell system (not all in 1 cell)</li>
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     <li>Gibson assembly of another combinatorial promoter using pKS001 backbone, lacI geneblock, ____ promoter geneblock, and sfGFP geneblock.</li>
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     <li>Gibson assembly of another combinatorial promoter using pKS001 backbone, lacI geneblock, B83 promoter geneblock, and sfGFP geneblock.</li>
     <li>Transformation of Gibson constructs into JM109 cells</li>
     <li>Transformation of Gibson constructs into JM109 cells</li>
     <li>Linear combinatorial constructs created via PCR of pAA001 (minipreps) to be tested in TXTL</li>
     <li>Linear combinatorial constructs created via PCR of pAA001 (minipreps) to be tested in TXTL</li>

Revision as of 17:27, 3 July 2014


WELCOME TO iGEM 2014!

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On this page you can document your project, introduce your team members, document your progress
and share your iGEM experience with the rest of the world!


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Notebook

Apparently there's a calendar feature to take care of this too???


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 Two

Monday, 6/23/14
  • Discussion of fallout from Friday's meeting, paper reading
  • Colony PCR and Minipreps of liquid cultures of colonies picked over the weekend
  • TXTL workshop starts today

Tuesday, 6/24/14
  • Discussed alternatives to current ComQXPA system, including 2-cell system (not all in 1 cell)
  • Gibson assembly of another combinatorial promoter using pKS001 backbone, lacI geneblock, B83 promoter geneblock, and sfGFP geneblock.
  • Transformation of Gibson constructs into JM109 cells
  • Linear combinatorial constructs created via PCR of pAA001 (minipreps) to be tested in TXTL

Wednesday, 6/25/14
  • Linear constructs and plasmid TXTL reactions of A90 promoter (pAA001) were set up and run
  • Gibson assembly of plasmid pAA002 containing B83 promoter
  • PCR of Gibson assemblies of pAA002 to create linear constructs to test in TXTL
  • TXTL reactions of B83 promoter (linear frag. from pAA002) were set up and run
  • Transformation of pAA002 Gibson assemblies into JM109

Thursday, 6/26/14
  • Analysis of yesterday's TXTL reactions (run overnight)
  • Colony PCR and miniprep of bacterial colonies transformed with pAA002. Minipreps shipped for sequencing.
 TXTL reaction data inconclusive. Most fluorescence signals are below negative control, and linear data frequently contradicts plasmid data with A90 promoter. No signal observed for B83 promoter.

Friday, 6/27/14
  • Continued to investigate different signalling pathways
  • Potential alternative pathways to investigate:
    • fsrABCD
    • agrBDCA
    • uhpABCT


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