Yeast Colony PCR: Difference between revisions

Revision as of 10:08, 17 January 2006

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 [[Silver: Colony PCR|Silver Lab: Yeast Colony PCR]]<br>
 [[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