IGEM:UNAM/2009/Notebook/IGEM project/2011/04/25

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How extract genomic DNA?

  • Today I learned how extract genomic DNA.

Method to extract genomic DNA

  • 1. Inoculate 5 ml of medium with Escherichia Coli DH5α and grow it for 12 hours.
  • 2. Put in the centrifugate the medium to 10 000 rpm for 1 minute.
  • 3. Resuspend the cell pill in 1 ml of TE 50:20, pH 8.0 and shake in the vortex.
  • 4. Put in the centrifugate for 1 minute and throw supernatant.
  • 5. Resuspend the cell pill in 0.4 ml of TE 50:20 and shake in the vortex.
  • 6. Put 50 μl of SDS to 10% and 50 μl of proteinase K to 2.5 mg/ml. Mix by inversion.
  • 7. Put alcohol 100% to the solution in a proportion 2:1, (2 ethanol: 1 solution).
  • 8. Quit the supernant with a syringe.
  • 9. Put alcohol 70% to the solution in a proportion 2:1, (2 ethanol: 1 solution).
  • 10.Quit the supernant with a syringe and dry with the DNA speed vac (5 minutes).
  • 11.Put 100 μl of TE-RNAsa (50μg/ml)and wait for 6 hours.