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

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson


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