Team:Paris Saclay/Notebook/August/22

Monday 22th 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.

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.