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 500 uL chilled 75% EtOH
   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]

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 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

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