Team:BCCS-Bristol/Protocols-Transformation using electroporation



Sterilization of the cuvettes:

 * 1) 	Open the cuvette and put both (lid and cuvette) into a UV light machine (CL-1000 Ultraviolet Crosslinker from UPV)
 * 2) 	Give 1200 “energy” (press “start” and wait for the end of the countdown)

Electroporation

 * 1) 	Prepare 950 μl sterile LB broth in a 1.5 ml tube and put in on ice
 * 2) 	Put the cuvettes at least 2 mins before adding the bacteria on ice
 * 3) 	Thaw the electric competent cells (E. coli DH5α) on ice (here 50 μl cells)
 * 4) 	Add the DNA to the cells (for good working DNA in high concentration like pUC19 with 10 pg/μl use 1 μl; for BioBrick DNA use 5 μl if it was prepared with 10 μl of water)
 * 5) 	Choose program Ec1 (BIORAD MicroPulser) and change the view to “ms” time
 * 6) 	Dry the wet cuvette with a paper towel and put it into the machine
 * 7) 	Give the pulse and notice the time: A pulse between 5-6 ms is a good value
 * 8) 	Add immediately 950 μl ice cold LB broth and transfer everything back into the tube
 * 9) 	Incubate for 1 h at 37°C 225 rpm
 * 10) 	Plate a part of the solution or spin the cells down, remove most of the supernatant (800-900 μl), resuspend them and plate all on a LB plate containing antibiotics
 * 11) 	Incubate the plates overnight at 37°C