Friday 5^st August

Lab work

Visualization

DNA Extraction of DS-TDcasN- and DS-SPcasN-

By Laetitia

The extraction was performed using the ChargeSwitc-Pro MiniPrep kit from Invitrogen. We followed this protocol.

The initial culture was at 20 mL so we increased (*4) the volumes until the precipitation.

Hence we used:

• 1 ml of resuspension buffer
• 1 ml lysis buffer
• 1 ml precipitation buffer

The colum was used several times in order to recover the maximum of DNA

DNA Extraction of DS-NMcasN- and pPS_001

By Mathilde

The extraction was performed on clone 11 for NM and clone 9 for 1.1 using the ChargeSwitc-Pro MiniPrep kit from Invitrogen. We followed this protocol.

Q5 PCR on pPS16_003, pPS16_004, pPS16_006 and pPS16_007

By Caroline

The PCR was carried out following the usual protocol adapted to 50µL and with a TM at 70°C. The specific primers for each parts were used. The products were put for migration on a 0.8%agarose gel with BET. Only, PCR form pPS16_003 and pPS16_004 showed a positive result.

PCR Clean-up with the NucleoSpin kit

By Caroline

The purification was carried out on PCR products 2.1, 2.2 and 4.1 following the usual protocol. But, I forgot to dilute the Buffer NT3 with 100mL ethanol before I began the purification and so there was no DNA observed on the 0.8% agarose gel.

File:T--Paris_Saclay--160722_Visualisation_PCRpPS16_002_7.JPG