Yeast Colony PCR

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See also: [[Silver: Colony PCR|Silver Lab: Yeast Colony PCR]]
See also: [[Silver: Colony PCR|Silver Lab: Yeast Colony PCR]]
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#Using sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolase
#Using sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolase

Revision as of 16:10, 21 September 2005

See also: Silver Lab: Yeast Colony PCR


  1. 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
  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 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
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