Team:USTC CHINA/Notebook/Protocols/Plasmid mini-prep

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<h1>Plasmid mini-prep</h1>
<p><span>Performed with Shanghai Sangon Plasmid miniprep kit.</span></br>
1. Preparation</br>
1) Make sure the RNase A has been added into Buffer P1;</br>
2) Make sure the ethanol has been added into Wash Solution;</br>
3) Make sure there is no deposit in Buffer P2 and P3;</br>
2. Collect the deposit of bacteria from 1.5ml~5ml bacteria liquid (centrifuge at 8,000×g for 2 min).</br>
3. Add 250μl Buffer P1 into the deposit and suspend it thoroughly.</br>
4. Add 250μl Buffer P2, mix gently and keep standing in room temperature for 2~4min.</br>
5. Add 350μl Buffer P3 and mix gently for 5~10 times.</br>
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).</br>
7. Add 500 μl Buffer DW1, centrifuge at 9,000×g for 30s</br>
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.</br>
9. Add 50~100μl Elution Buffer and collect the plasmid.</br></p>
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