Monday 22^th August

Lab work

Visualization

Low fidelity Dreamn Taq PCR of pPS16_016

By Mathilde

Clones 7,8,9 and 10 of pPS16_016 were amplified following the protocol :

• 1,5µL of Green Buffer
• 1µL of dNTPs
• 1µL of each primers IPS83 and IPS84
• 0,13µL of Dream Taq Polymerase
• 19,37 µL of H20

Colonies were put into sterile water, put through 5 minutes of denaturation before the addition of the mix. Annealing temperature was 57°c. After amplification, 1µL of each PCR product and 10µL of the DNA purple ladder were palced in wells and migrated at 100V for 30min.

Result of the PCR

Sequencing of 1.2, FKBP, FRB

Terrence and Léa

Was sent :

Purification on gel of gBlocks 3 and 4

Terrence and Mathilde

After amplification, 50µL of each PCR product diluted 6 tumes with 10 µL of the gel loadding dye, and 10µL of the DNA purple ladder were placed in wells and migrated at 100V for 30min. The extraction was carried out with the usual protocol.

Nano Drop :

Gel for the extraction

Purification of PSB1C3 with kit

Terrence

PSB1C3 was purified following the usual protocol. To be sure that PSB1C3 were linear, it was digested with DPN1.

Nano Drop :

Gibson assembly product transformation in DH5a cells

by Léa

Gibson assembly product (fragments 3 and 4 ligation) and control were transformed into DH5a cells, using 2 µL of DNA, and following the usual protocol.

pPS16_016 ligation product transformation in DH5a

By Léa

pPS16_016 ligation product of the 10/08/2016 was transformed into DH5a cells, using 2 µL of DNA, and following the usual protocol. Transformation products were plated on LB Agar chloramphenicol Petri dishes and incubated at 37°C ON.

Interlab Study

Transformation into DH5a cells

By Léa

Devices 2 and 3, cnegative control and positive control was transformed into DH5a cells, following the usual protocol. Transformation products were plated on LB Agar chloramphenicol Petri dishes and incubated at 37°C ON.

Device 1 culture

By Léa

Device 1 transformed DH5a glycerol stock was plated on LB Agar chloramphenicol Petri Dishes and incubated at 37°C ON.