Team:USTC CHINA/Notebook/Protocols/Gel Extraction

From 2013.igem.org

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Incubate at 75° C for 15-20 min or until gel melts completely.
Incubate at 75° C for 15-20 min or until gel melts completely.
Add 0.5 × Buffer DE-A volume of Buffer DE-B.</br>
Add 0.5 × Buffer DE-A volume of Buffer DE-B.</br>
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<img src="https://static.igem.org/mediawiki/2013/1/10/Gel_Extraction1.jpg" width="240" height="120" />  
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<img src="https://static.igem.org/mediawiki/2013/1/10/Gel_Extraction1.jpg" width="240" height="120" /> </br>
3. Binding sample DNA(Centrifuge at 5,000 - 6,000×g for 5 min).</br>
3. Binding sample DNA(Centrifuge at 5,000 - 6,000×g for 5 min).</br>
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<img src="https://static.igem.org/mediawiki/2013/0/0c/Gel_Extraction2.jpg" width="347" height="136" />   
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<img src="https://static.igem.org/mediawiki/2013/0/0c/Gel_Extraction2.jpg" width="347" height="136" />  </br>
4. Add 800 μl of Buffer W2 (5,000 - 6,000×g for 1 min).</br>
4. Add 800 μl of Buffer W2 (5,000 - 6,000×g for 1 min).</br>
Repeat wash with Buffer W2.</br>
Repeat wash with Buffer W2.</br>
Centrifuge empty plate for 5 min at 5,000 - 6,000×g to remove residue W2.</br>
Centrifuge empty plate for 5 min at 5,000 - 6,000×g to remove residue W2.</br>
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<img src="https://static.igem.org/mediawiki/2013/0/0c/Gel_Extraction3.jpg" width="347" height="136" />   
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<img src="https://static.igem.org/mediawiki/2013/0/0c/Gel_Extraction2.jpg" width="347" height="136" />  </br>
5. Elute with 50 μl of Eluent or Deionized Water.(5,000 - 6,000×g, 5min).</br>
5. Elute with 50 μl of Eluent or Deionized Water.(5,000 - 6,000×g, 5min).</br>
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<img src="https://static.igem.org/mediawiki/2013/6/69/Gel_Extraction4.jpg" width="349" height="93" />   
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<img src="https://static.igem.org/mediawiki/2013/6/69/Gel_Extraction4.jpg" width="349" height="93" />  </br>

Revision as of 16:25, 25 September 2013

Gel Extraction

Performed with AxyPrep 96-well DNA Gel Extraction Kit (type: AP-96-GX) Protocol 1. Excise gel slice containing DNA fragment of interest.
2. Add 3×sample volume of Buffer DE-A.
Incubate at 75° C for 15-20 min or until gel melts completely. Add 0.5 × Buffer DE-A volume of Buffer DE-B.

3. Binding sample DNA(Centrifuge at 5,000 - 6,000×g for 5 min).

4. Add 800 μl of Buffer W2 (5,000 - 6,000×g for 1 min).
Repeat wash with Buffer W2.
Centrifuge empty plate for 5 min at 5,000 - 6,000×g to remove residue W2.

5. Elute with 50 μl of Eluent or Deionized Water.(5,000 - 6,000×g, 5min).