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/revtrans">Reverse Transcriptase</a></td>
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/revtrans">Reverse Transcriptase</a></td>
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<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/mutagen">Mutagenisis</a></td>
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<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>
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<td>Content</td>
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<b>Digest</b></br></br>
<b>Digest</b></br></br>
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<p>
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1) Cast an agarose gel for purification of the cut product</br></br>
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1)Cast an agarose gel for purification of the cut product</br></br>
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2) Mix the digestion mixture in an eppendorf tube</br></br>
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2)Mix the digestion mixture in an eppendorf tube</br></br>
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3)Leave for 5 min. at 37°C (no shaking!)</br></br>
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3) Leave for 5 min. at 37°C (no shaking!)</br></br>
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4)Immidiately load the restriction mixture in the gel</br></br>
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4) Immidiately load the restriction mixture in the gel</br></br>
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5)Run the gel, cut out and purify bands of correct size.</br></br>
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5) Run the gel, cut out and purify bands of correct size.</br></br>
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Latest revision as of 21:09, 26 September 2012

iGEM TEAM ::: SDU-DENMARK courtesy of NIAID





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


Digest

1) 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


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