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Team:UNIPV-Pavia/Protocols/Precipitation
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
DNA precipitation with sodium acetate
(estimated time: 1 hour)
Materials needed:
- sodium acetate
- absolute ethanol
- ethanol 70%
- ddH2O
- previously cut plasmid
- Add 1/10 digestion volume of sodium acetate 3 M
- Add 2.5 digestion volume of absolute ethanol
- Freeze at -80°C for 30 min
- Centrifuge at 13000 rpm, 4°C for 20 min
- Decant supernatant
- Add 50 µl of 70% ethanol
- Centrifuge at 13000 rpm, 4°C for 20 min
- Remove all supernatant with a pipette
- Air dry pellet until ethanol is totally removed
- Elute with 5 µl ddH2O
