Daniel:Protocols/EtOHPrecip: Difference between revisions
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==Protocol== | ==Protocol== | ||
1. Add (X=volume sample): | 1. Add (X=volume sample): | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="background-color:#EEECE1;font-size:12pt;font-weight:bold" align="center" | |||
| width="65" height="16" | Chemical | |||
| width="65" | Volume | |||
|- style="background-color:#EEECE1;font-size:12pt" align="center" | |||
| height="16" | 100% EtOH | |||
| ≥ 2.5X | |||
|- style="background-color:#EEECE1;font-size:12pt" align="center" | |||
| height="16" | 3 M NaOAc | |||
| 0.1X | |||
|- style="background-color:#EEECE1;font-size:12pt" align="center" | |||
| height="16" | Glycoblue | |||
| 1/300X | |||
|} | |||
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 | For smaller fragments it is helpful to incubate overnight |
Revision as of 23:27, 17 April 2013
Ethanol Precipitation
Purpose: To purifiy DNA.
Protocol
1. Add (X=volume sample):
Chemical | Volume |
100% EtOH | ≥ 2.5X |
3 M NaOAc | 0.1X |
Glycoblue | 1/300X |
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