Team:USTC CHINA/Notebook/Protocols/Expression of proteins in Bacillus subtilis

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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)