User:University of Washington/14 August 2008

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==LuxR from AraC and TetR(Faifan)==
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<h4>QuikChange Approach</h4>
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- miniprepped QuikChange overnight cultures (rxn1 x 4, rxn2) and submit sequencing:Forward
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<h4>Elowitz</h4>
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-dilute 1:10 of digested BBa_Elowitz
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-3 way ligation(20 ul rxn)
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*mix
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<table border=1>
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<tr align="center">
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<th></th><th>dH20(ul)</th><th>Buffer(ul)</th><th>P1010-Kan(ul)</th><th>BBa_Elowitz(ul)</th><th>GFP(ul)</th><th>LuxR(ul)</th><th>T4 ligase(ul)</th>
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</tr>
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<tr align="center">
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<td>GFP</td><td>14.6</td><td>2</td><td>0.40</td><td>1</td><td>1</td><td>-</td><td>1>/td>
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</tr>
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<tr align="center">
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<td>LuxR</td><td>14.6</td><td>2</td><td>0.40</td><td>1</td><td>-</td><td>1</td><td>1>/td>
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</tr>
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</table>
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*30 mins incubate, room temp
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*10 mins denature, 65 degree Celsius
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*20 mins filtration
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-transformed into XL1-Blue and plated on Kan
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Back to [[Team:University_of_Washington/Notebook#Notebook]]
Back to [[Team:University_of_Washington/Notebook#Notebook]]

Latest revision as of 22:32, 14 August 2008

LuxR from AraC and TetR(Faifan)

QuikChange Approach

- miniprepped QuikChange overnight cultures (rxn1 x 4, rxn2) and submit sequencing:Forward

Elowitz

-dilute 1:10 of digested BBa_Elowitz

-3 way ligation(20 ul rxn)

  • mix
dH20(ul)Buffer(ul)P1010-Kan(ul)BBa_Elowitz(ul)GFP(ul)LuxR(ul)T4 ligase(ul)
GFP14.620.4011-1>/td>
LuxR14.620.401-11>/td>
  • 30 mins incubate, room temp
  • 10 mins denature, 65 degree Celsius
  • 20 mins filtration

-transformed into XL1-Blue and plated on Kan

Back to Team:University_of_Washington/Notebook#Notebook

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