Monday 29^th August

Lab work

Visualization

PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid

By Mahnaz

• 2.5 µL DreamTaq Buffer
• 0.5 µL of dNTPs (10mM)
• 1 µL of each primer (10µM) primer pJET R and F
• 0.13 μl of DreamTaq Pol
• 20 µl H2O

PCR was performed as follow:

Primers used were:

After amplification, 3 µL of each PCR products and 5 µL of DNA ladder were placed in wells and migrated at 100V during 30 min.

PCR products expected were :

Result of migration

The expected size was 474 pb and the 4 plasmids were sent to sequencing.

Séquençage

Samples preparation for sequencing

by Mahnaz

15 µL of plasmides plasmids coding FRB from colonies 4,5,9 and 12 were sent to be sequenced. 20 µL of the primers pJET F (5µM) and pJET R (5µM) were sent for sequencing.

NanoDrop Measurements

By Mahnaz