"
Team:NRP-UEA-Norwich/Protocol
From 2012.igem.org
Welcome to the NRP UEA iGEM 2012 Wiki Lab Book
Please choose the relevant link to access our diary of that week!
Week Zero | Week One | Week Two | Week Three | Week Four | Week Five | Week Six | Week Seven | Week Eight | Week Nine | Week Ten | Week Eleven | Lab Protocols | Experiments
1) Competent cells were thawed on ice for 15mins
2) Into pre-chilled 2ml tubes, 50 µL competent thawed cells and 2µL of re-suspended DNA were added and mixed through flicking.
3) These were put back into ice and left for 30mins.
4) From the ice, the tubes were taken and immersed in a 42 ºC water bath for 1 min.
5) After heat shocking, cells were incubated on ice for 5mins.
6) To these cells, 200 µL of pre-warmed LB broth (37 ºC) was added.
7) Cells were then incubated at 37 ºC for 2 hours and shaken every 5mins
8) From each tube two inoculations were made. 1) 20 µL of cells plus 180 µL of LB on chlorophenical LB agar plates, 2) 200 µL of cells. Agar plates contain chlorophenicol at a concentration of 50µg/ml.
9) These were incubated for 17 hours at 37 ºC.
