Difference between revisions of "Template:Team:Groningen/CONTENT/EXPERIMENTS/Competent E. Coli 1"

Line 17: Line 17:
  
 
<div class="text"> 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.
 
<div class="text"> 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.
</div>\
+
</div>
 
<div class="text"
 
<div class="text"
 
Used culture (OD: 0,24) for making competent cells (protocol Clement)  
 
Used culture (OD: 0,24) for making competent cells (protocol Clement)  

Revision as of 13:16, 6 August 2015

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.
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