Revision as of 13:43, 29 July 2010

Competent cells preparation

Inoculate 300 ml of LB broth with 1/20 volume of an overnight culture of the desired strain.
Grow the cell at 37°C (with shaking platform so as to mix the media equally amongst the cells)
Next Chill cells on ice and harvest by centrifugation at 4°C for 10 minutes
Resuspend the pellet in 100 ml of ice cold TFBII
Aliquot 200 µl volumes of the cell suspension into cooled sterile microfuge tubes and flash freeze in dry-ice
Store at -80°C

@@ Line 2: / Line 2: @@
 =Competent cells preparation=
+==Materials required==
+# 300ml of LB broth
+# 1/20 volume of an overnight culture
 ==Procedures==
-#Inoculate 300 ml with 1/20 volume of an overnight culture of the desired strain.
+# Inoculate 300 ml of LB broth with 1/20 volume of an overnight culture of the desired strain.
-#Grow the cell at 37°C (with shaking)
+# Grow the cell at 37°C (with shaking platform so as to mix the media equally amongst the cells)
-#Chill cells on ice and harvest by centrifugation at 4°C for 10 minutes
+# Next Chill cells on ice and harvest by centrifugation at 4°C for 10 minutes
-#Resuspend the pellet in 100ml of ice cold TFBII
+# Resuspend the pellet in 100 ml of ice cold TFBII
-#Aliquot 200 µl volumes of the cell suspension into cooled sterile microfuge tubes and flash freeze in dry-ice
+# Aliquot 200 µl volumes of the cell suspension into cooled sterile microfuge tubes and flash freeze in dry-ice
-#Store at -80°C
+# Store at -80°C
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