Yeast Colony PCR: Difference between revisions
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==Lab Protocols== | |||
[[Silver: Colony PCR|Silver Lab: Yeast Colony PCR]]<br> | |||
[[Endy:Yeast Colony PCR]] | |||
Revision as of 10:08, 17 January 2006
Lab Protocols
Silver Lab: Yeast Colony PCR
Endy:Yeast Colony PCR
- 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