Team:SDU-Denmark/labwork/Protocols/Checkdigest
From 2012.igem.org
(Difference between revisions)
Line 265: | Line 265: | ||
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/mutagen">Mutagenisis</a></td> | <td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/mutagen">Mutagenisis</a></td> | ||
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/pcrgelclean">PCR-,gel clean-up</a></td> | <td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/pcrgelclean">PCR-,gel clean-up</a></td> | ||
- | + | ||
</tr> | </tr> | ||
</table> | </table> |
Revision as of 17:04, 26 September 2012
mRNA Isolation | PCR | Miniprep | Check Digest |
3A-Assembly | Colony-PCR | Transformation | Gel-electrophoresis |
Reverse Transcriptase | Mutagenisis | PCR-,gel clean-up |
Check Digest
Digest mix, using Fastdigest enzymes
Digest1) Cast an agarose gel for purification of the cut product 2) Mix the digestion mixture in an eppendorf tube 3) Leave for 5 min. at 37°C (no shaking!) 4) Immidiately load the restriction mixture in the gel 5) Run the gel, cut out and purify bands of correct size. Digestion Mix
4 µL H2O 0,5 µL enzyme A 0,5 µL enzyme B 2 µL Fast Digest green buffer 3 µL dna product