Gel Extraction

Performed with Shanghai Sangon Plasmid miniprep kit. 1. Preparation 1) Make sure the RNase A has been added into Buffer P1; 2) Make sure the ethanol has been added into Wash Solution 3) Make sure there is no deposit in Buffer P2 and P3 2. Collect the deposit of bacteria from 1.5ml~5ml bacteria liquid (centrifuge at 8,000×g for 2 min) 3. Add 250μl Buffer P1 into the deposit and suspend it thoroughly 4. Add 250μl Buffer P2, mix gently and keep standing in room temperature for 2~4min 5. Add 350μl Buffer P3 and mix gently for 5~10 times 6. Precipitate the bacteria fragments (centrifuge at 12,000×g for 5~10 min) and extract the supernatant (centrifuge at 8,000×g for 30s) 7. Add 500 μl Buffer DW1, centrifuge at 9,000×g for 30s 8. Add 500 μl Wash Solution, centrifuge at 9,000×g for 30s. Repeat this process for one more time. Centrifuge the empty bottle at 9,000×g for 60s. 9. Add 50~100μl Elution Buffer and collect the plasmid.