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

 
(4 intermediate revisions by the same user not shown)
Line 39: Line 39:
 
|executor=Harm Ruesink
 
|executor=Harm Ruesink
 
|content=
 
|content=
−
<div class="text">Check transformation:
+
<div class="text">Checked transformation:
 
Transformation failed. White smear on 100% plate. Maybe the CM did not work properly.
 
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).</div>
 +
<div class="listing">
 +
<div class="item"> 1 µL of 50 pg/µL BBa_J04450 </div>
 +
</div>
 +
 +
<div class="text">
 +
Plated on LB agar + CM:
 +
</div>
 +
<div class="listing">
 +
<div class="item"> 10%  (10 µL) </div>
 +
<div class="item"> 90%  (90 µL) </div>
 +
</div>
 +
 +
<div class="text">
 +
Grown overnight in incubator at 37 °C.
 
</div>
 
</div>
 +
}}
  
 +
{{Team:Groningen/TEMPLATES/LOG
 +
|title=00:00, 09 July 2015 - 00:00, 09 July 2015
 +
|executor=Harm Ruesink
 +
|content=<div class="text">Checked transformation: No colonies on 10% plate. A white smear on 90% plate. No red colonies, so transformation failed.</div>
 
}}
 
}}

Latest revision as of 10:10, 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
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.

Harm Ruesink
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.
Harm Ruesink