Endy:Yeast Colony PCR

 

Method

1. Using sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolyase
   • 3 uL of 1 U/mL Zymolyase stock solution
   • 47 uL of water
2. Incubate cell suspension at 37 C for 30 minutes.
3. Incubate cell suspension at 95 C for 10 minutes.
4. Use 5 uL of the cell suspension for a 50 uL PCR reaction

Zymolase Stock Solution

• 10,000 U Zymolyase (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