Team:USTC CHINA/Notebook/Protocols/Gel Extraction

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<p>Performed with AxyPrep 96-well DNA Gel Extraction Kit (type: AP-96-GX)
<p>Performed with AxyPrep 96-well DNA Gel Extraction Kit (type: AP-96-GX)
Protocol
Protocol
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1. Excise gel slice containing DNA fragment of interest.
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1. Excise gel slice containing DNA fragment of interest.</br>
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2. Add 3×sample volume of Buffer DE-A.
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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.
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Add 0.5 × Buffer DE-A volume of Buffer DE-B.
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Add 0.5 × Buffer DE-A volume of Buffer DE-B.</br>
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<img src="https://static.igem.org/mediawiki/2013/4/4d/2013igemustc-china_Protocol1.png" width="580" height="200" />  
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3. Binding sample DNA(Centrifuge at 5,000 - 6,000×g for 5 min)
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3. Binding sample DNA(Centrifuge at 5,000 - 6,000×g for 5 min)</br>
   
   
4. Add 800 μl of Buffer W2 (5,000 - 6,000×g for 1 min)
4. Add 800 μl of Buffer W2 (5,000 - 6,000×g for 1 min)
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Repeat wash with Buffer W2
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Repeat wash with Buffer W2</br>
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Centrifuge empty plate for 5 min at 5,000 - 6,000×g to remove residue W2.
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Centrifuge empty plate for 5 min at 5,000 - 6,000×g to remove residue W2.</br>
   
   
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5. Elute with 50 μl of Eluent or Deionized Water.(5,000 - 6,000×g, 5min)
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5. Elute with 50 μl of Eluent or Deionized Water.(5,000 - 6,000×g, 5min)</br>
   
   
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Revision as of 16:11, 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)

notebook

Retrieved from "http://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols/Gel_Extraction"