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Team:USTC CHINA/Notebook/Protocols/Gel Extraction
From 2013.igem.org
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Protocol | Protocol | ||
1. Excise gel slice containing DNA fragment of interest.</br> | 1. Excise gel slice containing DNA fragment of interest.</br> | ||
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2. Add 3×sample volume of Buffer DE-A.</br> | 2. Add 3×sample volume of Buffer DE-A.</br> | ||
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> | ||
| - | + | <img src="https://static.igem.org/mediawiki/2013/1/10/Gel_Extraction1.jpg" width="240" height="120" /> | |
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> | ||
| - | + | <img src="https://static.igem.org/mediawiki/2013/0/0c/Gel_Extraction2.jpg" width="347" height="136" /> | |
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> | ||
| - | + | <img src="https://static.igem.org/mediawiki/2013/0/0c/Gel_Extraction3.jpg" width="347" height="136" /> | |
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> | ||
| - | + | <img src="https://static.igem.org/mediawiki/2013/6/69/Gel_Extraction4.jpg" width="349" height="93" /> | |
Revision as of 16:24, 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).
