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.

Harm

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.

Harm

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.

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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

Harm