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User:University of Washington/11 July 2008
From 2008.igem.org
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- Selected 6 colonies each from two 2-ul-DNA plates and inoculated onto two new plates (each divided into 6 regions) to get a single colony of each. (?) Left plates in the cabinet for two days growth. | - Selected 6 colonies each from two 2-ul-DNA plates and inoculated onto two new plates (each divided into 6 regions) to get a single colony of each. (?) Left plates in the cabinet for two days growth. | ||
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== Lambda Red Recombineering of RP4 (Bryan) == | == Lambda Red Recombineering of RP4 (Bryan) == | ||
| - | Repeated yesterday's PCR amplification of recombinant fragment. Included correct primer concentrations and reaction buffer this time. For template DNA, used RP4, pAC88, and yEP13, all vectors carrying a TetR cassette. PCR failed anyway. Putative source of error: template plasmids carry a tetR cassette that is not homologous to our primers. | + | Repeated yesterday's PCR amplification of recombinant fragment. Included correct primer concentrations and reaction buffer this time. Ran parallel reactions with primers designed both by myself and Scott. For template DNA, used RP4, pAC88, and yEP13, all vectors carrying a TetR cassette. PCR failed anyway. Putative source of error: template plasmids carry a tetR cassette that is not homologous to our primers. |
Designed additional primers for future Lambda Red experiments. | Designed additional primers for future Lambda Red experiments. | ||
Revision as of 20:12, 18 July 2008
LuxR from AraC and TetR
- Transformations of mutated AraC plasmid into XL1 Blue succeed!! Got many colonies in both of 2ul DNA plates, few in 15 ul DNA plates, none in negative control.
- Selected 6 colonies each from two 2-ul-DNA plates and inoculated onto two new plates (each divided into 6 regions) to get a single colony of each. (?) Left plates in the cabinet for two days growth.
Lambda Red Recombineering of RP4 (Bryan)
Repeated yesterday's PCR amplification of recombinant fragment. Included correct primer concentrations and reaction buffer this time. Ran parallel reactions with primers designed both by myself and Scott. For template DNA, used RP4, pAC88, and yEP13, all vectors carrying a TetR cassette. PCR failed anyway. Putative source of error: template plasmids carry a tetR cassette that is not homologous to our primers.
Designed additional primers for future Lambda Red experiments.
AHL production in yeast (Bryan)
After yesterday's transformation of C0161 failed, making the third try using a new approach, concluded the part is faulty. Attempted transformation of related part C0061 LuxI-LVA into Top10.
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