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User:University of Washington/12 August 2008
From 2008.igem.org
LuxR from AraC and TetR(Faifan)
-minipreped and submit sequencing for QuikChange(trail#3)
-Results for QuikChange(trail#4)looked good. Colonies were streaked out in new Amp plate.
- 4 colonies in reaction1 plate
- 1 colony in reaction2 plate
- none in negative(-primers)
- many in positive(-Dpn1)
-sequencing results of the promoters (PCR from Elowitz's) on vector pSB1AC3 came out half good.
- Ligation at room temp seemed to yield better products. Sequences from 4 minipreps matched what we wanted ==> the promoter is successfully biobricked!!!
- Ligation overnight at 4 degree Celsius was not as effective? One miniprep out of four showed to have promoter between the prefix and suffix. The rest still contained P1010(death gene) which was weird.
-Two out of four colonies from the old marks on 25 degree Celsius ligation plate were selected and overnight with Tsy + Amp.
AHL Expression in Yeast (Bryan)
Ligated C0161 LuxI BioBrick into YEp426. Ran three reactions at 10:1, 6:1, and 2:1 insert:plasmid ratio.
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