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Team:UNIPV-Pavia/Protocols/PlasmidExtraction
From 2008.igem.org
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The protocols we used
- LB medium preparation
- Plasmid resuspension from IGEM paper spots
- Transformation
- Plasmid extraction
- BioBrick digestion with restriction enzymes
- DNA gel extraction
- DNA precipitation with sodium acetate
- Antarctic Phosphatase
- Ligation
- PCR
Plasmid extraction
(Estimated time: 3 hours if you use QIAGEN Plasmid Mini Kit; 1 hour if you use QIAGEN QIAprep Spin Miniprep Kit)
Materials needed:
- 5 ml or 9 ml (for higher yields) overnight mini-culture
- QIAGEN Plasmid Mini Kit or QIAGEN QIAprep Spin Miniprep Kit
- Follow QIAGEN standard protocol: [http://www1.qiagen.com/literature/render.aspx?id=369&tp=4 QIAGEN Plasmid Mini Kit protocol] or [http://www1.qiagen.com/literature/render.aspx?id=370&tp=4 QIAGEN QIAprep Spin Miniprep protocol]
- We used QIAGEN Plasmid Mini Kit in the first 2 weeks of our activity to perform miniprep, when we didn't have QIAGEN QIAprep Spin Miniprep Kit at our disposal. Then, we always used QIAGEN QIAprep Spin Miniprep Kit for its higher yield.
