Daniel:Protocols/EtOHPrecip: Difference between revisions
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=Ethanol Precipitation= | =Ethanol Precipitation= | ||
[[Daniel Jacobsen|Back to Main]] | |||
Purpose: To purifiy DNA. | Purpose: To purifiy DNA. | ||
Line 27: | Line 29: | ||
For smaller fragments it is helpful to incubate overnight | For smaller fragments it is helpful to incubate overnight | ||
3. Centrifuge at 4 C | 3. Centrifuge at 4 C | ||
4. Remove supernatant and add | 4. Remove supernatant and add 750 uL chilled 75% EtOH | ||
5. Centrifuge at minutes at 4 C | 5. Centrifuge at minutes at 4 C | ||
6. Dry the pellet in the hood | 6. Dry the pellet in the hood |
Latest revision as of 18:33, 2 July 2014
Ethanol Precipitation[edit]
Purpose: To purifiy DNA.
Protocol[edit]
1. Add:
Chemical | Volume |
100% EtOH | ≥ 2.5X |
3 M NaOAc | 0.1X |
Glycoblue | 1/300X |
where X = volume of the sample
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 750 uL chilled 75% EtOH 5. Centrifuge at minutes at 4 C 6. Dry the pellet in the hood 7. Resuspend pellet in desired volume ddH20 8. Store DNA at -20C
Recommended Centrifuge Times[edit]
Spin | Size Tube | Time | RPM |
1 | 15 mL | 30 | 3000 |
1 | 1.5 mL | 25 | 12000 |
2 | 1.5 mL | 10 | 10000 |