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.

Friday, 6/27/14

• Continued to investigate different signalling pathways
• Potential alternative pathways to investigate:
  • fsrABCD
  • agrBDCA
  • uhpABCT