Team:UNIPV-Pavia/Protocols/Precipitation
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(New page: <h1>DNA precipitation with sodium acetate</h1> ''(estimated time: 1 hour)'' <br> <br> '''Materials needed:''' *'''sodium acetate''' *'''absolute ethanol''' *'''ethanol 70%''' *'''ddH2O'''...) |
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<h1>DNA precipitation with sodium acetate</h1> | <h1>DNA precipitation with sodium acetate</h1> | ||
''(estimated time: 1 hour)'' | ''(estimated time: 1 hour)'' |
Revision as of 17:26, 22 June 2009
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DNA precipitation with sodium acetate
(estimated time: 1 hour)
Materials needed:
- sodium acetate
- absolute ethanol
- ethanol 70%
- ddH2O
- previously cut plasmid
- We use it only if the miniprep did not give us a high yield of DNA
- 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