User:University of Washington/6 August 2008

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LuxR from AraC and TetR(Faifan)

-Tranformation of ligation product at room temp(P1010 on pSB1AC3 and promoter) succeed? There were too much growth, so the culture was streaked out in new Amp plate.

-Continued ligation process (the one at 4 degree Celcius overnight): denatured enzyme, filterd, tranformed into XL1-Blue, and grew on Amp plate.

-Discovered today that the dsDNA added for previous QuikChange reaction was way excessive due to unit misunderstanding.

-Did first part of QuikChange (set up reaction + theymocycling overnight)

  • nanodropped R0080(AraC) = 113.5 ng/ul, 1.9-260/280, 2.19-260/230
  • Elongation step 68 degree Celcius for 7 mins was removed in theymocycling.

MG1655Z1(Faifan)

-Got growth in Amp plate from MG1655Z1 stock. Have to find out what plasmid contains Amp resistance from literature.

-Got growth in Amp and a bit in Tet plates from MG1655Z1 Tsy#2 plate. This said that there were still some unknown plasmids in the strain. Two colonies from section 3 and 5 were picked and grew overnight in Tsy.



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