Endy:Yeast Colony PCR

Back to Yeast Colony PCR

Method

 * 1) Using sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolyase
 * 2) *3 uL of 1 U/mL Zymolyase stock solution
 * 3) *47 uL of water
 * 4) Incubate cell suspension at 37 C for 30 minutes.
 * 5) Incubate cell suspension at 95 C for 10 minutes.
 * 6) 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

Related Topics
Silver: Colony PCR