Endy:Yeast Colony PCR

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''Back to [[Yeast Colony PCR]]''
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==Method==
#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
#*3 uL of 1 U/mL Zymolase stock solution
#*3 uL of 1 U/mL Zymolase stock solution
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*Add H2O to 10 mL
*Add H2O to 10 mL
*Store at -20 C
*Store at -20 C
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==Related Topics==
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[[Silver: Colony PCR]]

Revision as of 13:10, 17 January 2006

Back to Yeast Colony PCR

Method

  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

Related Topics

Silver: Colony PCR

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