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Team:Paris-Saclay/Project/Notebook/Week 10
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Revision as of 22:24, 26 September 2012
Week 10
6th August
- Sending of the B1 sample to sequencing with two pairs of primers
- K-274100 Forward and Reverse
- Plasmid pSB1A2 Forward and Reverse
7th August
- Liquid culture of B1 in order to prepare a glycerol stock
- Receipt of new primers
- 2 Forward
- 3 Reverse
- Plasmid Reverse
8th August
| Amplification of the plasmid pSB1A2, BBa_K274100 and BBa_K115017 by PCR with the new primers. Visualization by electrophoresis on a 2% Agarose gel for BBa_K115017 and a 0.8% Agarose gel for BBa_K274100 and the plasmid pSB1A2. We are expecting a band at 147 bp for BBa_K115017, 3408 bp for BBa_K274100 and 2079 for the plasmid. |  |
PCR program used:
9th August
| New amplification by PCR of the plasmid pSB1A2, BBa_K274100 and BBa_K115017 as it has been done the day before. |  |
- Digestion by EcoRI of the plasmid that contains BBa_K274100.
10th August
| Amplification by PCR of BBa_K274100 already digested by EcoRI. Visualization by electrophoresis on a 0.8% Agarose gel. |  |
PCR program used:
| Miniprep of BBa_K274100 followed by Nanovue to determine the concentration of the sample |  |
| Miniprep of the Plasmid pSB1A2 followed by Nanovue to determine the concentration of the sample |  |
- Digestion by HindIII of the plasmid pSB1A2 in order to linearize it.
| Digestion of BBa_K115017 by DPNI to eliminate the plasmid matrix. Visualization by electrophoresis on a 2% Agarose gel. We are expecting a band at 123 bp. |  |
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