S. pombe Colony PCR

For E. coli colony screening, see Colony PCR. For full S. pombe genomic DNA extraction, see S. pombe Genomic DNA Midiprep (zymolyase method).

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 zymolyase (10mg/ml) and incubate at 37°C for 1h (zymolyase 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.