Team:Groningen/Protocols

From 2010.igem.org

(Difference between revisions)
(New page: CaCl2 Competent Cells '''Day 1:''' -Inoculate 5 ml LB( TY) medium with ''E.coli'' from glycerol stock -Grow overnight at 37 oC '''Day 2:''' -Inoculate 20 ml LB medium with 200 ...)
Line 3: Line 3:
'''Day 1:'''
'''Day 1:'''
-
-Inoculate 5 ml LB( TY)  medium with ''E.coli'' from glycerol stock
+
-Inoculate 5 ml [[LB( TY)  medium]] with ''E.coli'' from glycerol stock
-Grow overnight at 37 oC  
-Grow overnight at 37 oC  
Line 15: Line 15:
-Spin down 5 minute at 4000 rpm at 4 oC
-Spin down 5 minute at 4000 rpm at 4 oC
-
-Resuspend in 10 ml chilled 0,1 M caCl2 (from here, keep on ice! )
+
-Resuspend in 10 ml chilled 0,1 M CaCl2 (from here, keep on ice! )
-Inoculate on ice for 20 minutes
-Inoculate on ice for 20 minutes

Revision as of 09:53, 24 August 2010

CaCl2 Competent Cells

Day 1:

-Inoculate 5 ml LB( TY) medium with E.coli from glycerol stock

-Grow overnight at 37 oC

Day 2:

-Inoculate 20 ml LB medium with 200 μl overnight culture

-Grow at 37 oC until OD600 = 0,3-0,5(+/- 2 hours)

-Spin down 5 minute at 4000 rpm at 4 oC

-Resuspend in 10 ml chilled 0,1 M CaCl2 (from here, keep on ice! )

-Inoculate on ice for 20 minutes

-Spin down as before

-Remove supernatant

-Resuspend in 2 ml 0,1 M CaCl2 /10% glycerol

-Divide 100 μl aliquots

-Store competent cells in - 80 oC