Difference between revisions of "Template:Team:Groningen/CONTENT/EXPERIMENTS/Competent E. coli 2"

Line 34: Line 34:
 
Grown overnight in incubator at 37 °C.</div>
 
Grown overnight in incubator at 37 °C.</div>
 
}}
 
}}
 +
 +
{{Team:Groningen/TEMPLATES/LOG
 +
|title=00:00, 08 July 2015 - 00:00, 08 July 2015
 +
|executor=Harm Ruesink
 +
|content=
 +
<div class="text">Check transformation:
 +
Transformation failed. White smear on 100% plate. Maybe the CM did not work properly.
 +
</div>

Revision as of 07:56, 7 August 2015

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
Harm Ruesink
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.
Harm Ruesink

{{Team:Groningen/TEMPLATES/LOG |title=00:00, 08 July 2015 - 00:00, 08 July 2015 |executor=Harm Ruesink |content=

Check transformation:

Transformation failed. White smear on 100% plate. Maybe the CM did not work properly.