Endy:Yeast Colony PCR - Revision history

Reshma P. Shetty at 13:02, 10 September 2009

2009-09-10T13:02:06Z

←Older revision		Revision as of 13:02, 10 September 2009
Line 18:		Line 18:
	==Related Topics==		==Related Topics==
	[[Silver: Colony PCR]]		[[Silver: Colony PCR]]
		+
		+	[[Category:Yeast]] [[Category:Protocol]]

Ssutton at 20:03, 16 February 2006

2006-02-16T20:03:37Z

←Older revision		Revision as of 20:03, 16 February 2006
Line 1:		Line 1:
	''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 ~~Zymolase~~	+	#Using sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolyase
-	#*3 uL of 1 U/mL ~~Zymolase~~ stock solution	+	#*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.
Line 9:		Line 9:

	'''Zymolase Stock Solution'''		'''Zymolase Stock Solution'''
-	*10,000 U ~~lyticase~~ (Sigma)	+	*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

Barry Canton at 17:10, 17 January 2006

2006-01-17T17:10:06Z

←Older revision		Revision as of 17:10, 17 January 2006
Line 1:		Line 1:
		+	''Back to [[Yeast Colony PCR]]''
		+	==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
Line 13:		Line 15:
	*Add H2O to 10 mL		*Add H2O to 10 mL
	*Store at -20 C		*Store at -20 C
		+
		+	==Related Topics==
		+	[[Silver: Colony PCR]]

Barry Canton at 17:08, 17 January 2006

2006-01-17T17:08:49Z

New page

#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