Team:Hawaii/Notebook/2008-10- 6

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Things we did today

Wetlab work

Construction of secretion device (cont.)

Grace
  • PCR'd nir, nir+rbs, rbs+GFP+tt#2, plac+rbs, slr1+GFPf
  • Verified PCR products on 0.8% agarose gel
  • Overnight RE digest:
  • EcoRI/SpeI:
  • nir
  • nir+rbs
  • plac+rbs
  • XbaI/PstI;
  • slr1+GFPf
  • pilA+GFPf+tt
  • rbs+GFP+tt#2
  • HindIII/BamHI:
  • J33207

Drylab Work

Sequencing analysis

Grace
  • Correct:
  • rbs+GFP+tt #1, 2
  • BBpRL1383a:
  • All four sequenced colonies had a G->C single bp substitution at position 229 of part (in CAP binding site)
  • Colony 18 confirmed (best quality sequence)
  • Colony 1 confirmed (abiguities at 225, 583, 591, 552 of part)
  • Colony 3 (abiguities at 530, 513 of part)
  • Colony 13 (low quality sequence, but BioBrick MCS is present)
  • Incorrect:
  • nir+rbs+GFP #4, 7
  • nir+rbs+slr1+GFPf #8
  • No sequence obtained:
  • plac+rbs
  • nir+rbs+GFP #1
  • nir+rbs+slr1+GFPf #2
  • nir+rbs+pilA #17


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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