Tuesday 9^th August

Lab work

Visualization

Heat shock transformation

By Charlène and Terrence

For 1.2, 4.2, SgNm, SgSt1, Detection, FRB and FKBP gBlocks, the results of the sequencing were not as expected.So, we transformed pPS16_002, pPS16_008, pPS16_010, pPS16_011, pPS16_012, pPS16_013 and pPS16_014 in DH5a. We followed the usual protocol. 50µL of cells were streaked on LB + Amp (50µg/mL) + IPTG + xGal.

Phusion PCR on pPS16_006 and pPS16_007

By Caroline

The PCR was carried out following the usual protocol adapted to 50µL and with a TM at 72°C. The specific primers for each parts were using. The products were put to migrated on a 0.8%agarose gel with BET.

pUC19 digestion with HincII

By Alice 5 µL of pUC19 plasmids were digested with 5µL of tango buffer 10X, 38µL of sterile water, and 1 µL of HincII enzyme. The mix was incubated at 37°C for 1 hour. After incubation, 1µL of HincII enzyme was added again, and the mix was incubated 1 hour again. Digetion products were migrated on a gel. We expected to have a band of 2.7 Kb.

pUC19 ligation with gBlocks ATG linker RFB, detection, St sgRNA, ATG linker FKBP, Nm sgRNA, 1.2 and 4.2

By Alice