Revision as of 09:59, 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

@@ Line 119: / Line 119: @@
 Finally put it in a incubator at 37°C for 30 minutes
-==== Transformation of pPS3 in DH5α ====
-We add 100 µl of DH5α in the pPS3 ligation tube and we follow the classic [https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_competent_e.coli_by_CaCl2 protocol] of transformation.
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