Template:Team:Groningen/CONTENT/EXPERIMENTS/Competent E. coli 2
00:00, 06 July 2015 - 00:00, 06 July 2015
A colony was picked from a plate (± 14 days old, stored in fridge) and grown overnight in liquid media (5 mL LB in 50 mL tube) in shaker incubator at 37 °C, 200 rpm
00:00, 07 July 2015 - 00:00, 07 July 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, 200 rpm for 2 hours.
Used culture 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:
100% (100 µL)
Grown overnight in incubator at 37 °C.
00:00, 08 July 2015 - 00:00, 08 July 2015
Checked transformation:
Transformation failed. White smear on 100% plate. Maybe the CM did not work properly.
Tested competent cells again with Transformation efficiency kit (BBa_J04450, 50 pg/µL). Protocol: Heat Shock transformation (protocolbook). 1 µL of 50 pg/µL BBa_J04450
Plated on LB agar + CM:
10% (10 µL)
90% (90 µL)
Grown overnight in incubator at 37 °C.
00:00, 09 July 2015 - 00:00, 09 July 2015
Checked transformation: No colonies on 10% plate. A white smear on 90% plate. No red colonies, so transformation failed.