From 2009.igem.org
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| - | Writing Paper
| + | Transformation of week-long Ligation of promoter library and vector pCS26 |
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| - | * Finished writing modeling paper that summarizes what we will be doing once the circuit is completed. The experiments should not take long to accomplished and these results can be tabulated in Matlab quickly. | + | * Promoter Library product (from primer extension PCR) was ligated with pCS26 vector for 9 days with T4 Invitrogen Ligase at room temperature (~21<sup>o</sup>C). This product was transformed into both XL Gold Ultracompetent and Top 10 cells. |
| - | * 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.
| + | * Grew these cells on LB plates with Kan resistance at 37<sup>o</sup>C for 16 hours. |
| - | * 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.
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| - | * 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 02:31, 29 July 2009
University of Calgary
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CAROL
Transformation of week-long Ligation of promoter library and vector pCS26
- Promoter Library product (from primer extension PCR) was ligated with pCS26 vector for 9 days with T4 Invitrogen Ligase at room temperature (~21oC). This product was transformed into both XL Gold Ultracompetent and Top 10 cells.
- Grew these cells on LB plates with Kan resistance at 37oC for 16 hours.
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CHINMOYEE
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EMILY
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FAHD
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IMAN
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JAMIE
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JEREMY
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KATIE
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KEVIN
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MANDY
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PATRICK
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PRIMA
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STEFAN
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VICKI
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