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User:Andriana/18 June 2012
From 2012.igem.org
Contents |
I. 15-16, 29-30 mutations at 61C melting temperature
II. 9-10 and retried 7-8 at 55C melting temperature
III. 21-22 at 67C melting temperature
IV. Meeting with Prof. Isaacs
V. Observed and recorded results of the purple experiment:
| Plate | Properties |
|---|---|
| 2 A | 150 colonies |
| 2 B | 400 colonies |
| 2 C | 430 colonies |
| 2 D | 900 colonies |
| 4 A | 18 colonies |
| 4 B | 30 colonies |
| 4 C | 300 colonies |
| 4 D | 800 colonies |
- Analysis
- Freezing at -80C does increase E. coli death rates
- Greatest E. coli death was observed at .1M NaCl, so optimal concentration for cell death is less than 1M but more than .01M NaCl.
- All things said, all data points to the fact that freezing in salt solutions does not provide an effective selection pressure for finding mutants with the most effective AFP or that produce large amounts of AFP because the normal, untransformed control cells exhibited greater survival rates than their transformed counterparts
- The purple experiment showed that antibiotic plates alone did not cause the reduced colony numbers in the red experiment because the trend of lesser survival of cells 3 and 4 also persisted in the purple experiment
VI. Digested PCR reactions with DPN1:
- Add 1 microliter of DPN1 for 50microliters of reaction
- Incubate solution at 37C for an hour
- Heat kill DPN1 enzymes at 80C for 20 minutes
