Daniel:Protocols/EtOHPrecip: Difference between revisions
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1/300X glycoblue (co-precipitate, helps you see the DNA) | 1/300X glycoblue (co-precipitate, helps you see the DNA) | ||
2. Incubate for at least 1 hour at -80 C | 2. Incubate for at least 1 hour at -80 C | ||
For smaller fragments it is helpful to incubate overnight | |||
3. Centrifuge | 3. Centrifuge at 4 C | ||
4. Remove supernatant and add 500 uL chilled 75% EtOH | |||
5. Spin at 10000 rpm for 10 minutes at 4 C | |||
6. Dry the pellet in the hood | |||
7. Resuspend pellet in desired volume ddH20 | |||
8. Store DNA at -20C |
Revision as of 23:25, 17 April 2013
Ethanol Precipitation
Purpose: To purifiy DNA.
Protocol
1. Add (X=volume sample): At least 2.5X 100% Ethanol 0.1X 3M NaOAc 1/300X glycoblue (co-precipitate, helps you see the DNA) 2. Incubate for at least 1 hour at -80 C For smaller fragments it is helpful to incubate overnight 3. Centrifuge at 4 C 4. Remove supernatant and add 500 uL chilled 75% EtOH 5. Spin at 10000 rpm for 10 minutes at 4 C 6. Dry the pellet in the hood 7. Resuspend pellet in desired volume ddH20 8. Store DNA at -20C