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Endy:Yeast Colony PCR: Difference between revisions
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''Back to [[Yeast Colony PCR]]'' | ''Back to [[Yeast Colony PCR]]'' | ||
==Method== | ==Method== | ||
#Using sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml | #Using sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolyase | ||
#*3 uL of 1 U/mL | #*3 uL of 1 U/mL Zymolyase stock solution | ||
#*47 uL of water | #*47 uL of water | ||
#Incubate cell suspension at 37 C for 30 minutes. | #Incubate cell suspension at 37 C for 30 minutes. | ||
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'''Zymolase Stock Solution''' | '''Zymolase Stock Solution''' | ||
*10,000 U | *10,000 U Zymolyase (Sigma) | ||
*1 mL KHPO4 buffer, pH 7.5 (adjusted with KOH) | *1 mL KHPO4 buffer, pH 7.5 (adjusted with KOH) | ||
*0.2 mL 5M NaCl | *0.2 mL 5M NaCl | ||
