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Team:Calgary/26 July 2009
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* Plated the week-long ligation of the promoter library in both XL Gold Ultra competent cells and Top 10 cells, but there is no growth. | * Plated the week-long ligation of the promoter library in both XL Gold Ultra competent cells and Top 10 cells, but there is no growth. | ||
* A reason that we thought of that might have caused minimal colonies is the phosphatase step. The 32 primers that we have synthesized does not have 5' phosphates. If we phosphatase the vector, that might result in no ligation. | * A reason that we thought of that might have caused minimal colonies is the phosphatase step. The 32 primers that we have synthesized does not have 5' phosphates. If we phosphatase the vector, that might result in no ligation. | ||
| - | * Re-started restriction enzyme digestion on vector pCS26 with XhoI and BamHI at 37<sup>o</ | + | * Re-started restriction enzyme digestion on vector pCS26 with XhoI and BamHI at 37<sup>o</sup>C for 4 hours. Then I ligated the promoter with the vector with Quick Ligase for 5 minutes. Then I transformed the plasmids into both XL Gold Ultracompetent cells and Top 10 cells. Incubate at 37<sup>o</sup>C for 16 hours. |
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Revision as of 02:14, 29 July 2009
UNIVERSITY OF CALGARY
