Latest revision as of 18:55, 16 September 2008

Cloning of EnvZ*

Defroze of the cells on ice
Add 5 µL of DNA in 100 µL of competent cells (ligation products or pUC19 for positive control)
30 min on ice
Heat schock 45sec at 42°C
2 min on ice
Add 900 µL of SOC
Incubate 1h at 37°C
Centrifugate 5 min at 6000 rpm
Remove 800 µL
Plate the 200 µL left on LB + amplicilline
Incubate O/N at 37°C

@@ Line 54: / Line 54: @@
 |3,4 µL
 |}
-*10 min at room temperature, followed by 7 hours at about 16°C.
+*10 min at room temperature
+*followed by 7 hours at about 16°C.
 ==Transformation of E. coli DH5 alpha competent cells==
-*defrosing of the cells on ice
+* Defroze of the cells on ice
-*adding of 5 µL of ligation products in 100 µL of competent cells in 14 mL falcon polypropylene tubes
+* Add 5 µL of DNA in 100 µL of competent cells (ligation products or pUC19 for positive control)
-*30 min on ice
+* 30 min on ice
-*thermal schock of 45 s at 42°C
+* Heat schock 45sec at 42°C
-*2 min on ice
+* 2 min on ice
-*adding of 900 µL of SOC
+* Add 900 µL of SOC
-*1 hour at 37°C
+* Incubate 1h at 37°C
-*contrifugation 5 min 6000 rpm
+* Centrifugate 5 min at 6000 rpm
-*removal of 800 µL
+* Remove 800 µL
-*plating the 200 µL left on LB + amplicilline
+* Plate the 200 µL left on LB + amplicilline
-*overnight incubation at 37°C
+* Incubate O/N at 37°C