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Team:UNIPV-Pavia/Protocols/PlasmidExtraction
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Plasmid extraction
QIAGEN Plasmid Mini Kit
(estimated time: 3 hours)
Materials needed:
- Overnight mini-culture
- QIAGEN Plasmid Mini Kit
- You can find the detailed protocol at: [http://www1.qiagen/HB/QIAGENPlasmidPurification_EN http://www1.qiagen/HB/QIAGENPlasmidPurification_EN]
- Incubate on ice for 30-45 min.
- Heat shock: 42°C for 1 min.
- Put transformed TOP10 tube on ice and then add 250 µl SOC medium.
- Incubate 2 hours at 37°C, 220 rpm.
- Centrifuge 10 min, 1200 rpm.
- Remove 150 ul of bacteria-free supernatant.
- Plate the remaining part of solution (resuspending the bacteria) on a proper agar plate.
- Incubate overnight at 37°C.
