Team:Groningen/Notebook/Competent e coli 1
Blue Bio Energy
Competent E. Coli 1
Competent E. coli cells were made.
To make competent E. coli cells.
A few red colonies on the 90% plate (± 22 colonies).
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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 (5 mL LB in 50 mL tube) in shaking incubator at 37 °C, 220 rpm.
00:00, 30 April 2015 - 00:00, 30 April 2015
Transferred overnight culture in fresh LB (pre-warmed). 250 µL culture in 25 mL LB (1/100 dilution) and grown in shaker incubator at 37 °C, 220 rpm for 2 hours.
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).
1 µL of 50 pg/µL BBa_J04450
Plate on LB agar + CM:
10% (10 µL)
90% (90 µL)
control
00:00, 1 Juni 2015 - 00:00, 1 Juni 2015
Checked transformation:
A few red colonies on the 90% plate (± 22 colonies).