Isopropanol Precipitation of DNA 

Materials

  1. DNA solution to be concentrated (from Phenol/CIA separation)
  2. 10M NH4Ac
  3. Sterile microcentrifuge tubes
  4. Isopropanol, Molecular biology grade
  5. 80% 200-proof ethanol
  6. Bechtop Centrifuge
  7. Speed-Vac
  8. Pipet and tips
  9. Thermoshaker (optional)
Procedure
  1. Take 200µl DNA solution to 1.5ml microcentrifuge tube.
  2. Add 10M NH4Ac to 1.5M final (35.3µl) and mix
    • Avoid shearing DNA, Vortex speed depond on DNA base length.
    • Too much NH4Ac may cause co-precipitation of primers.
  3. Add 0.6 volume of isopropanol (141.2µl) and mix.
  4. Incubate at RT for 15 min.
  5. Spin at top speed (14,000 rpm) at RT for 15 min and remove supernatant by asperation.
  6. Wash with 1 DNA volume of 80% ethanol.
  7. Vacum dry.
  8. Dissolve pellet in T108.0 by pipetting (and heating at 50oC if neccesary).
  9. Store sample at 4oC.  

Note

  • Iisopropanol precipitation can also remove primers from PCR reactions but ethanol precipitation will not.

 

 

 

 

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