Friday 22nd August

Lab work

D - Lemon Scent

pPSI digestion

pPSI dephosphorylation

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)

Ligation of LS/GS/PS in pPSI

For each synthase, put:

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

Transformation of pPS3 in DH5 alpha

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

Back to the calendar