Tuesday 16^th August

Lab work

Visualization

2.1-2.2 and 3.1-3.2 ligation

By Charlène

8µL of 2.1 purify PCR products, 8µL of 2.2 purify PCR products, 2µL of ligase T4 Buffer and 2L of ligase T4 were mixed. 8µL of 3.1 purified PCR products, 8µL of 3.2 purified PCR products, 2µL of ligase T4 Buffer and 2µL of ligase T4 were mixed. They were incubated for 1h at RT.

Q5 PCR on the ligation products and pPS16_008 clones 1 and 2

By Charlène

The PCR was carried out with a new protocol: Q5 PCR recipe

Steps for PCR :

An specific primers for each part was used.

The products were migrated on a 0.8% agarose gel with BET.

Gel Electrophoresis of PCR products fragments 2, 3 and gblock 4.1.JPG

All the bands have the good size so purifications will be done on PCR products.

PCR Clean-up with the NucleoSpin kit

By Mahnaz

The purification was carried out on PCR product PSB1C3 (linearized) and 4.2 also the ligation products PS16003-PS16004 and PS16005-PS16006 following the usual protocol.

4.1-4.2 ligation

By Mahnaz

8 µL of 4.1 purified PCR product, 8µL of 4.2 purified PCR product, 2 µL of ligase T4 Buffer and 2 µL of ligase T4 were mixed. The ligation reaction mixture were incubated overnight at 4°C.