Revision as of 09:50, 25 August 2014

Friday 22nd August

After 2 hours, add 1.5µL of Alkaline Phosphatase (AP) and let it 1 hour.

Then, inactivation of AP at 65°C during 15 minutes.

We've made a electrophoresis to check the digestion.

(photo)

And let it in the freezer for 3 hours.

After 3 hours, take out the ligation tubes and add 100 µl of competent DH5α in each tube.

Then proceed to the transformation protocol.

- 20 minutes at 4°C

- 2 minutes 30 seconds at 42°C

- 2 minutes at 4 °C

- Add 0.9 ml of LB into the tubes

Finally put it in a incubator at 37°C for 30 minutes

We add 100 µl of DH5 alpha in the pPS3 ligation tube and we follow the classic protocol of transformation.

@@ Line 104: / Line 104: @@
 And let it in the freezer for 3 hours.
-====Transformation in competent E.coli DH5 Alpha====
+====Transformation in competent E.coli DH5α====
-After 3 hours, take out the ligation tubes and add 100 µL of competent DH5 Alpha in each tube.
+After 3 hours, take out the ligation tubes and add 100 µl of competent DH5α in each tube.
-Then procede to the transformation protocol.
+Then proceed to the transformation protocol.
 - 20 minutes at 4°C
@@ Line 116: / Line 116: @@
 - 2 minutes at 4 °C
-- Add 0.9 mL of LB into the tubes
+- Add 0.9 ml of LB into the tubes
-And finally put it in a incubator at 37°C during 30 minutes
+Finally put it in a incubator at 37°C for 30 minutes
 ==== Transformation of pPS3 in DH5 alpha ====