Team:Ryerson Toronto/Notebook
Notebook
Week 1
P. areuginosa, strain mPA01, grown on LB agar plate overnight at 37°C
Gentamycin, at a final concentration of 30μg/mL-100μg/mL, was used to counter select
Single colony grown overnight in LB broth
Genomic DNA extracted
LB broth was spun down, resuspended in PBS, and boiled for 20 min. The supernatant was then transferred to a new tube
Week 2
PCR
samples were prepared in GC and HF Phusion buffer master mixes, with the following sample dilutions: no dilution, 1:50, 1:100, 1:250, 1:1000
Gel Electrophoresis
agarose gel, at a final concentration of 0.8% made in TBE, was run at 100V for 60min
Week 3
Transformation
For restriction digestion for transformation, the following enzymes were used: kpni and xhol
Samples from Transformation were applied to a gel
1% agarose gel was made with TAE, and run at 40V for 60min
T4 ligation
