Daniel:Protocols/EtOHPrecip: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Djacobse
No edit summary
>Djacobse
No edit summary
Line 5: Line 5:
==Protocol==
==Protocol==
   1. Add (X=volume sample):
   1. Add (X=volume sample):
    At least 2.5X 100% Ethanol
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
    0.1X 3M NaOAc
|- style="background-color:#EEECE1;font-size:12pt;font-weight:bold" align="center"
    1/300X glycoblue (co-precipitate, helps you see the DNA)
| 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