S. pombe Colony PCR

Contents

1. Procedure

1. Procedure

1. Resuspend medium sized colony in 5μl of sterile H2O in 1.5ml eppendorf tube.
2. Add 2 μl of zymolase (10mg/ml) and incubate at 37°C for 1h (zymolase from Seikagaku - cat#: 120493).
3. Add 10 μl of 0.02N NaOH.
4. Incubate 5 min at 95°C, then immediately 5 min on ice.
5. Quick-spin, vortex thoroughly and use 2 μl for a 50 μl PCR reaction.