Daniel:Protocols/EtOHPrecip: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Djacobse
No edit summary
>Djacobse
No edit summary
Line 9: Line 9:
     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
      For smaller fragments it is helpful to incubate overnight
   3. Centrifuge on
   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