User:University of Washington/12 August 2008

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==LuxR from AraC and TetR(Faifan)==
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-minipreped and submit sequencing for QuikChange(trail#3)
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-Results for QuikChange(trail#4)looked good. Colonies were streaked out in new Amp plate.
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*4 colonies in reaction1 plate
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*1 colony in reaction2 plate
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*none in negative(-primers)
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*many in positive(-Dpn1)
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-sequencing results of the promoters (PCR from Elowitz's) on vector pSB1AC3 came out half good.
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*Ligation at room temp seemed to yield better products. Sequences from 4 minipreps matched what we wanted ==> the promoter is successfully biobricked!!!
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*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.
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-Two out of four colonies from the old marks on 25 degree Celsius ligation plate were selected and overnight with Tsy + Amp.
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==AHL Expression in Yeast (Bryan)==
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Ligated C0161 LuxI BioBrick into YEp426.  Ran three reactions at 10:1, 6:1, and 2:1 insert:plasmid ratio.
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Back to [[Team:University_of_Washington/Notebook#Notebook]]
Back to [[Team:University_of_Washington/Notebook#Notebook]]

Latest revision as of 19:03, 15 August 2008

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.

Back to Team:University_of_Washington/Notebook#Notebook

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