Difference between revisions of "Yeast Colony PCR"

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See also: [[Silver: Colony PCR|Silver Lab: Yeast Colony PCR]]
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''Back to [[Protocols]]''
  
#Using sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolase
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==Lab Protocols==
#*3 uL of 1 U/mL Zymolase stock solution
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*[[Silver: Colony PCR|Silver Lab: Yeast Colony PCR]]
#*47 uL of water
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*[[Endy:Yeast Colony PCR]]
#Incubate cell suspension at 37 C for 30 minutes.
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*[[Blackburn:Yeast_Colony_PCR_v2.0|Blackburn Lab:  Quick and Easy Yeast Colony PCR v2.0]]
#Incubate cell suspension at 95 C for 10 minutes.
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*[[Smolke:Colony_PCR_tsb|Smolke Lab: Colony PCR (Travis' version)]]
#Use 5 uL of the cell suspension for a 50 uL PCR reaction
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*[[BE.109:Bio-material_engineering/PCR_of_gold_binding_candidates| 20.109 lab protocol, steps 2 and 4]]
  
'''Zymolase Stock Solution'''
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[[Category:Protocol]]
*10,000 U lyticase (Sigma)
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[[Category:Yeast]]
*1 mL KHPO4 buffer, pH 7.5 (adjusted with KOH)
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[[Category:DNA]]
*0.2 mL 5M NaCl
 
*5 mL 100% glycerol
 
*Add H2O to 10 mL
 
*Store at -20 C
 

Latest revision as of 14:59, 20 March 2009

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