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Team:UNIPV-Pavia/Protocols/PlasmidExtraction
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Contents |
The protocols we used
- LB medium preparation
- Plasmid resuspension from IGEM paper spots
- Transformation
- Plasmid extraction
- BioBrick digestion with restriction enzymes
- DNA gel extraction
- Antarctic Phosphatase
- Ligation
- PCR
Plasmid extraction
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.
QIAGEN Plasmid Mini Kit
(estimated time: 3 hours)
Materials needed:
- 5 ml or 9 ml (for higher yields) overnight mini-culture
- QIAGEN Plasmid Mini Kit
- Followed QIAGEN standard protocol: [http://www1.qiagen.com/literature/render.aspx?id=369&tp=4 QIAGEN Plasmid Mini Kit protocol]
QIAGEN QIAprep Spin Miniprep Kit
(estimated time: 1 hour)
Materials needed:
- 5 ml or 9 ml (for higher yields) overnight mini-culture
- QIAGEN QIAprep Spin Miniprep Kit
- We followed standard protocol: [http://www1.qiagen.com/literature/render.aspx?id=370&tp=4 QIAGEN QIAprep Spin Miniprep Kit]
