Team:Hawaii/Notebook/2008-08-22

From 2008.igem.org

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Things we did today

Wetlab work

Verification of transformants

EtBr stained 2% agarose gel ran at 95V for 1.5 hours. Five microliters of PCR reaction were loaded into each lane.
Grace
Construct Colony forming units
nir+rbs 187
plac+rbs 0
rbs+GFP 1
GFP+tt 0
  • Colony PCR of transformants
  • Ran PCR reaction on 2% agarose gel
  • Can't tell 15bp difference between GFP and rbs+GFP
  • Restreaked single colony and inoculated for plasmid prep to confirm
  • nir+rbs colonies #11-19 appear promising (larger than nir)
  • Restreaked colony #19 and inoculated for plasmid prep to confirm

Plasmid prep of J33207

Grace

Overnight RE digest

Grace
  • BB-pRL1383a and J33207 with XbaI and PstI
  • pSB1A2 with EcoRI and PstI
  • GFP, GFPf, nir with EcoRI and SpeI

Made LB+amp100 plates

Grace

Ligation

Margaret

  • ligation of rep+pSB1A3, P1lytic region+pSB1A3, aada(BB&pRL1383a)+pSB1A3

Omega interposon

Margaret
  • streak on Sm&Sp plate to confirm insertion.

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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