Endy:Yeast Colony PCR
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- Using sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolase
- 3 uL of 1 U/mL Zymolase stock solution
- 47 uL of water
- Incubate cell suspension at 37 C for 30 minutes.
- Incubate cell suspension at 95 C for 10 minutes.
- Use 5 uL of the cell suspension for a 50 uL PCR reaction
Zymolase Stock Solution
- 10,000 U lyticase (Sigma)
- 1 mL KHPO4 buffer, pH 7.5 (adjusted with KOH)
- 0.2 mL 5M NaCl
- 5 mL 100% glycerol
- Add H2O to 10 mL
- Store at -20 C