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 :

Result of the purification

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 :