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Star Republic: Guide for Biologists

PCR amplification of yeast clones

Virtualab Protocols (Chang Bioscience)
Zymolyase

1. Dilute yeast culture 1:5 or 1:10. Add 1-2 ml of to a PCR reaction mix.

2. Add to each 50 ml PCR reaction mix 0.5 ml Zymolyase (2.5 units).

3. Incubate at 37 C for 5 minutes.

4. Proceed to PCR reactions.

Boiling

1. Pick a single clone or 1-2 ml of yeast culture.

2. Add to PCR reaction mix without enzyme.

3. Boil for 5 minutes and spin briefly.

4. Add Taq DNA polymerase.

5. Proceed to PCR reactions.

SDS

1. Pick a single clone using a toothpick.

2. Add to 20 ml 0.25% SDS.

3. Vortex and centrifuge.

4. Transfer 1 ml to PCR reaction mix.

5. Proceed to PCR reactions.