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.

Legend

1. pPSI+Apra digested via PacI, dephosphorylated 5µl
2. pPSI+Apra digested via PacI, dephosphorylated 5µl
3. ladder 5µl

Ligation of BBa_K517003, BBa_K762100, and p cola in pPSI

And let it in the freezer for 3 hours.

Transformation in competent E.coli DH5α

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

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