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User:University of Washington/24 July 2008
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[[UW Glycerol Stocks| Glycerol stock]] of pUB307 (RP1 conjugation plasmid) | [[UW Glycerol Stocks| Glycerol stock]] of pUB307 (RP1 conjugation plasmid) | ||
| - | [[ | + | [[Team:University_of_Washington/Protocols#Conjugation| bacteria-yeast conjugation protocol #2]]<br> |
·pUB307 + pAC88<br> | ·pUB307 + pAC88<br> | ||
·pAC88 | ·pAC88 | ||
Latest revision as of 17:37, 30 July 2008
Contents |
LuxR from AraC and TetR
- miniprepped mutated AraC (reaction1 x 4) (reaction2 x 4) in 30 ul EB.
- ordered sequencing for mutated AraC with forward primers.
- ran gel for R0080 (miniprepped yesterday) to see if AraC DNA in 6 tubes had same length. All except tube two appeared to be around 2 kb(as expected), so they were combined into one.
LuxR from pLac
-Cells transformed with R0010 and E0420 digest successfully grew on amp plate. Overnight culture started for sequencing.
-Overnight culture of DH5a+lacIq strain started. Needed for future transformations.
Lambda Red Recombineering of RP4 (Bryan)
Optimization of Lambda Red PCR protocol continued. Reduced annealing temperature to 40C. Ran parallel reactions to compare effect of dNTP concentration. Amplification occurred at 0.4 mM but not at 1 mM--the concentration indicated in our original protocol. Protocol will be revised. Additional benefit of reduced annealing temperature is inconclusive.
AHL Expression in Yeast (Bryan)
XS double digest of yEP426-GPD, yEP426-ADH, and C0161.
RP1 conjugation
Glycerol stock of pUB307 (RP1 conjugation plasmid)
bacteria-yeast conjugation protocol #2
·pUB307 + pAC88
·pAC88
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