Team:Hawaii/Notebook/2008-09- 5

From 2008.igem.org

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Things we did today

Wetlab work

Construction of Broad-Host-Range Vector Intermediates

Margaret
  • Plasmid Prep: Everything that didn't work yesterday
  • 2 Plac + B0030, 1 Plac + B0034, aadA(BB)colony 5, pSB3k3, pSB1A3
  • Restriction Digest: same as Yesterday
  • Ligation
  • Transformation into DH5alpha (we need more competent cells)

Verification of transformants from yesterday

EtBr stained 2% agarose gel ran at 83V for 1.75 hours. Two microliters of PCR product were loaded into each well.
Grace
Construct Colonies
plac+rbs (no vector) 0
C0012 derived vector 12
C0012 derived vector, SAP treated 163
3A: plac+rbs+vector >300
  • BBpRL1338a related plates all had a lawn (no distinct colonies observed). Too low antibiotic conc.? Will attempt to restreak/replate on Mon.
  • Colony PCR of 3A assembly related colonies
  • Oops. Loaded 1 μl colony #11 into colony #10's well
  • Colony #10 has potential. Will restreak on Mon.

Drylab Work

Sequencing

Margaret
  • Analysis of Sequencing submitted (oriV1-4, 2 aadA(BB), 2 aadA (pRL))
Grace
  • Confirmed:
  • nir+rbs
  • rbs+GFP
YAY!

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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