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User:University of Washington/23 July 2008
From 2008.igem.org
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| - | ==Lambda Red Recombineering of RP4 == | + | ==Lambda Red Recombineering of RP4 (Bryan) == |
Troubleshooting of PCR continued. Attempted PCR on control template at 1.5 mM, 2 mM, 2.5 mM, and 3 mM magnesium chloride. No amplicon at 1.5 mM, which conflicts with results from yesterday's PCR. Best amplicon appeared at 2 mM. Since the results are inconsistent, it is difficult to optimize the reaction concentration based on this experiment. | Troubleshooting of PCR continued. Attempted PCR on control template at 1.5 mM, 2 mM, 2.5 mM, and 3 mM magnesium chloride. No amplicon at 1.5 mM, which conflicts with results from yesterday's PCR. Best amplicon appeared at 2 mM. Since the results are inconsistent, it is difficult to optimize the reaction concentration based on this experiment. | ||
Revision as of 22:53, 23 July 2008
LuxR from AraC and TetR
- Miniprepped AraC R0080
- Results from Transformation of mutated AraC
- Positive control : a lot of growth
- Negative control : some growth (expected NO growth because primers weren't added.)
- Reaction one : some growth
- Reaction two : some growth
- Picked 4 colonies from each reaction and prepared overnight culture.
- Primers for Elowitz's promoter were designed and ordered.
Lambda Red Recombineering of RP4 (Bryan)
Troubleshooting of PCR continued. Attempted PCR on control template at 1.5 mM, 2 mM, 2.5 mM, and 3 mM magnesium chloride. No amplicon at 1.5 mM, which conflicts with results from yesterday's PCR. Best amplicon appeared at 2 mM. Since the results are inconsistent, it is difficult to optimize the reaction concentration based on this experiment.
Mini-prep of pSIM5 had no DNA in eluate. Putative error: pSIM5 does not have cam cassette leading to loss of plasmid or failure to grow in ON culture. Will not repeat mini-prep since pSIM5/host is in glycerol stock and not necessary for immediate plans.
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