Team:SDU-Denmark/labwork/Protocols/Checkdigest
From 2012.igem.org
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<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/PCR">PCR</a></td> | <td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/PCR">PCR</a></td> | ||
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/Miniprep">Miniprep</a></td> | <td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/Miniprep">Miniprep</a></td> | ||
- | <td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/Checkdigest">Check Digest</a></td> | + | <td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/Checkdigest"><b>Check Digest</b></a></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<tr> | <tr> | ||
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/revtrans">Reverse Transcriptase</a></td> | <td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/revtrans">Reverse Transcriptase</a></td> | ||
- | <td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/mutagen"> | + | <td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/mutagen">Mutagenesis</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> | ||
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<h1> Check Digest </h1> | <h1> Check Digest </h1> | ||
- | <h2> </h2> </b> | + | <p> |
+ | <h2>Digest mix, using Fastdigest enzymes</h2> | ||
+ | </br> | ||
+ | |||
+ | <b>Digest</b></br></br> | ||
+ | <p> | ||
+ | 1) Cast an agarose gel for purification of the cut product</br></br> | ||
+ | |||
+ | 2) Mix the digestion mixture in an eppendorf tube</br></br> | ||
+ | |||
+ | 3) Leave for 5 min. at 37°C (no shaking!)</br></br> | ||
+ | |||
+ | 4) Immidiately load the restriction mixture in the gel</br></br> | ||
+ | 5) Run the gel, cut out and purify bands of correct size.</br></br> | ||
+ | </br> | ||
+ | <b>Digestion Mix</b></br> | ||
+ | <pre> | ||
+ | <pre> | ||
+ | 4 µL H2O | ||
+ | 0,5 µL enzyme A | ||
+ | 0,5 µL enzyme B | ||
+ | 2 µL Fast Digest green buffer | ||
+ | 3 µL dna product | ||
+ | </pre> | ||
+ | </pre> | ||
+ | </br> | ||
+ | <p> | ||
+ | </br> | ||
Latest revision as of 21:09, 26 September 2012
mRNA Isolation | PCR | Miniprep | Check Digest |
3A-Assembly | Colony-PCR | Transformation | Gel-electrophoresis |
Reverse Transcriptase | Mutagenesis | 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