Revision as of 14:06, 22 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)

For each synthase, put:

And let it in the freezer during 3 hours.

After 3 hours, take out the ligation tubes and add 100 µL of competent DH5 Alpha in each tube.

Then procede 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

And finally put it in a incubator at 37°C during 30 minutes

@@ Line 61: / Line 61: @@
 And let it in the freezer during 3 hours.
+====Transformation in competent E.coli DH5 Alpha====
+After 3 hours, take out the ligation tubes and add 100 µL of competent DH5 Alpha in each tube.
+Then procede 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
+And finally put it in a incubator at 37°C during 30 minutes
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