Monday 18^th July

Lab work

Preparation of LB solid and liquid stock

By Caroline and Terrence

- 1L of LB liquid : 20g/L powder LB + 1L water μQ
- 500mL of LB solid : 500mL of LB liquid + 7.5g/L of Agar
The all was put in the autoclave for sterilization.

Getting DNA closer

Electrophoresis of PCR products DS_SPcasN, DS_TDcasN, pZA21 and pZA31

By Caroline

PCR products obtained the 13/07/2016 were put to migrate for 30min in a 0,8% agarose gel.

Biobrick characterization

LB culture of electro-transformed BL21 cells (Cf)

By Mathilde

Each transformation : pclTAA + K1372001, pcl_TAG + K1372001 and pcl_Tq + K1372001 was added to 3mL of LB and one or two antibiotics.

Three conditions were tested for each transformation :

• 15µL of Chloramphenicol (30µg/mL)
• 5 µL of Streptomycin (50µg/mL)
• 15µL of Chloramphenicol (30µg/mL) + 5 µL of Streptomycin (50µg/mL)

Cultures were put in incubation at 37°c, 180 rpm.

Visualization

Electrophoresis of PCR products pPS16_004

By Caroline

PCR products obtained the 13/07/2016 were put to migrate for 30min in a 0,8% agarose gel

Transformation of DH5α with 4.1 and 2.2

By Alice and Terrence

As the result of PCRs on clone 2.2 and 4.1 didn't gave us expected results. We supposed that bacteria used for the PCR didn't integrate the plasmids. So we made another transformation of Dh5α competent cells.

We made a dilution with the rest of the ligation product with 5μl of sterile H₂O and we follow the protocol of heat shock competent cells transformation.

Finally, we spread cells on Petri dishes (LB + Amp 50μg/mL + xGal 0.25μL/mL + IPTG0 0.1 μL/mL) in duplicate .