Difference between revisions of "Template:Team:Groningen/CONTENT/LOGBOOK/Competent e coli 2"
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|goal=<html>To make competent E. coli cells.</html> | |goal=<html>To make competent E. coli cells.</html> | ||
| − | |description=<html> | + | |description=<html>It was attempted to make competent E. coli cells. |
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|conclusion=<html>No red colonies were visible, so the transformation failed and no competent E. coli cells were made. | |conclusion=<html>No red colonies were visible, so the transformation failed and no competent E. coli cells were made. | ||
Latest revision as of 10:21, 7 August 2015
Competent E. coli 2
It was attempted to make competent E. coli cells.
To make competent E. coli cells.
No red colonies were visible, so the transformation failed and no competent E. coli cells were made.
<a class="postscriptum protocol" href="https://2015.igem.org/Team:Groningen/Protocols_and_Protocols/No_protocol">Finibus sapien, nec cursus...</a>
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.