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EPF-Lausanne/4 August 2008
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| + | == Cell culture and miniprep == | ||
| + | |||
| + | The DNA which was obtained on 01/08 by miniprep was put on a gel. The gel was clean and correct for the DNA with AmpR, but somehow unneat for the KanR/AmpRKanR ones. One which had been in very high concentration (300 ng/uL) seemed overloaded. The others seemed to contain genomic DNA, which could come from contamination or could be due to the use of spin columns from a different miniprep kit. We will in any case grow them out again with different Kan stock and proceed tomorrow with a digestion reaction, of shorter than overnight, to check the authenticity of the parts. | ||
| + | |||
| + | == PCR == | ||
| + | |||
| + | A PCR with the following primer-DNA sets was done : | ||
| + | |||
| + | <table width="207" border="1"> | ||
| + | <tr> | ||
| + | <td width="128">Primer</td> | ||
| + | <td width="63"><div align="center">Biobrick</div></td> | ||
| + | <td width="63"><div align="center">RhlR</div></td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>DNA</td> | ||
| + | <td><div align="center">RhlR (I0466)</div></td> | ||
| + | <td><div align="center">RFP (E1010)</div></td> | ||
| + | </tr> | ||
| + | |||
| + | </table> | ||
| + | |||
| + | The negative control (RFP with RhlR primer) did not work. The two first positives with Biobrick primers worked. However, the positive with RhlR primers failed. But this RhlR sequence was reported to be inconsistent, which could account for our trouble. | ||
Revision as of 13:00, 4 August 2008
Cell culture and miniprep
The DNA which was obtained on 01/08 by miniprep was put on a gel. The gel was clean and correct for the DNA with AmpR, but somehow unneat for the KanR/AmpRKanR ones. One which had been in very high concentration (300 ng/uL) seemed overloaded. The others seemed to contain genomic DNA, which could come from contamination or could be due to the use of spin columns from a different miniprep kit. We will in any case grow them out again with different Kan stock and proceed tomorrow with a digestion reaction, of shorter than overnight, to check the authenticity of the parts.
PCR
A PCR with the following primer-DNA sets was done :
| Primer | Biobrick |
RhlR |
| DNA | RhlR (I0466) |
RFP (E1010) |
The negative control (RFP with RhlR primer) did not work. The two first positives with Biobrick primers worked. However, the positive with RhlR primers failed. But this RhlR sequence was reported to be inconsistent, which could account for our trouble.
