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Team:UNIPV-Pavia/Protocols/AntarcticPhosphatase
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
Antarctic Phosphatase
(estimated time: 1 hour and 30 min)
Materials needed:
- NEB Antarctic Phosphatase
- 10X NEB Antarctic Phosphatase buffer
- Cut and gel-extracted vector
- Add the proper amount of 10X buffer to a final concentration of 1X (e.g. 2 µl of 10X buffer in a final volume of 20 µl).
- Add 1 µl of Antarctic Phosphatase (up to 5 µg of cut vector).
- Incubate at 37°C for 1 hour (Antarctic Phosphatase works).
- Incubate at 65°C for 15 min (Antarctic Phosphatase inactivation).
