Crude Genomic DNA Lysate for PCR

Crude Genomic DNA Lysate

1. Add isolated colony (or 2 uL broth culture) to 40 uL of sterile nuclease free H2O in a PCR tube.
2. Run thermocycler program "Lysate"
             55C for 10 minutes
             80C for 10 minutes
             20C for infinite
3. Add 80 uL of sterile nuclease free H2O to the PCR tube.
4. Centrifuge tubes on PCR centrifuge (Gibbs Lab, Teaching Lab) at 6000 rpm for 5 minutes to pellet.
5. Store lysates at -20C. Lysates still viable for a few months.

Use 1-2 uL of lysate for PCR reactions.

Note: The pellet will be cellular debris and should be avoided. The supernatant contains the crude genomic DNA.


//EWW