User:University of Washington/26 August 2008

From 2008.igem.org

(Difference between revisions)
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==LuxR from AraC and TetR(Faifan)==
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<h4>ligation from gel extraction</h4>
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-cultures of plasmid construct transformation ended up growing on Kan plates after incubating overnight at 36 degree Celsius
 +
 +
-selected 3 colonies from each plate(LuxR, GFP) and grew overnight to do sequencing
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 +
<h4>new ligation with CIP reaction</h4>
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-miniprep I0462 x 3, ran gel and combined tubes
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 +
-restriction digest GFP and LuxR as in the table from yesterday(8/25)
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-PCR purified digested promoter
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-nanodropped the purified promoter: 261.9 ng/ul, 1.87(260/280), 2.04(260/230)
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-did CIP reaction (Ingrid)
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*mix 10X CIP Buffer + 0.5 units of CIP per ul/DNA + DNA(have final concentration of 0.5 ul/10 ul)
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*incubated 37 degree Celsius 1 hour
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*stored at -20
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Back to [[Team:University_of_Washington/Notebook#Notebook]]
Back to [[Team:University_of_Washington/Notebook#Notebook]]

Revision as of 20:22, 26 August 2008

LuxR from AraC and TetR(Faifan)

ligation from gel extraction

-cultures of plasmid construct transformation ended up growing on Kan plates after incubating overnight at 36 degree Celsius

-selected 3 colonies from each plate(LuxR, GFP) and grew overnight to do sequencing

new ligation with CIP reaction

-miniprep I0462 x 3, ran gel and combined tubes

-restriction digest GFP and LuxR as in the table from yesterday(8/25)

-PCR purified digested promoter

-nanodropped the purified promoter: 261.9 ng/ul, 1.87(260/280), 2.04(260/230)

-did CIP reaction (Ingrid)

  • mix 10X CIP Buffer + 0.5 units of CIP per ul/DNA + DNA(have final concentration of 0.5 ul/10 ul)
  • incubated 37 degree Celsius 1 hour
  • stored at -20

Back to Team:University_of_Washington/Notebook#Notebook

Retrieved from "http://2008.igem.org/User:University_of_Washington/26_August_2008"