Team:Hawaii/Notebook/2008-07-17

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Things we did today

Wetlab work

PCR

Grace
PCR products on an EtBr stained 4% agarose gel run at 119V for 27 min then 95V for 25 min. Ten microliters of PCR reactions were loaded into each well.
  • Colony PCR for GFP fusion brick and new pRL1383a to verify inserts
  • Sequencing (25 mu;l reactions)
  • nir promoter
  • slr2016
  • pilA
  • BBa_B0015
  • BBa_B0024
  • BBa_B0030
  • BBa_C0012
  • BBa_E0040
  • BBa_J04430
  • BBa_J33207
  • BBa_R001
  • Ran all PCR products on a 4% gel to check
  • Gel ran funny -- loading dye ran 2x as fast as the DNA itself
  • 119V too much to handle? Wavy bands
  • Poor ladder resolution on gel
  • No bands seen for GFP fusion, pRL1383a, R0010
  • Incorrect bands for B0015 (multiple bands, what are the two bands at the top?) and J33207 (size too small, stuff still in wells)
  • Will redo plasmid preps for R0010, B0015, J33207
  • Will redo PCR reaction tomorrow for pRL1383a, GFP fusion, B0015, J33207, R0010

Plasmid Prep

Grace
  • Inoculated 7 ml LB + amp100 with single colony of B0015, R0010, J04430 for redo of plasmid prep tomorrow

Restreak to Purify

Grace
  • Restreaked GFP fusion and pRL1383a (MCS replaced) transformants to purify

pSB3K3 plasmid prep

Margaret


Discussion

Quote of the Day

At the team meeting today:

"We have cookies, we can start."


Ten cookies later...


"The more cookies I have, the better I think!"


- Dr. Callahan


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