Template:Team:Groningen/CONTENT/EXPERIMENTS/Competent E. Coli 1
00:00, 28 April 2015 - 00:00, 28 April 2015
Plated E. coli DH5 alpha on LB agar and grew at 37°C for 24h.
00:00, 29 April 2015 - 00:00, 29 April 2015
Picked a colony from LB agar and grew overnight in liquid media (5mL LB in 50mL tube) in shaking incubator at 37°C, 220rpm.
00:00, 30 April 2015 - 00:00, 30 April 2015
Transferred overnight culture in fresh LB (pre-warmed). 250µL culture in 25mL LB (1/100 dilution) and grew in shaker incubator at 37°C, 220rpm for 2h.
Used culture (OD: 0,24) for making competent cells (protocol Clement)
Tested competence of competent E. coli with Transformation efficiency kit (BBa_J04450, 50 pg/µL). Protocol: Heat Shock transformation (protocolbook).
1µL of 50pg/µL BBa_J04450
Plate on LB agar + CM:
10% (10µL)
90% (90µL)
control
00:00, 1 Juni 2015 - 00:00, 1 Juni 2015
Checked Transformation:
A few red colonies on the 90% plate (± 22 colonies).