Electroporation Protocol ( C2986 )
Prepare 17mm x 100mm round - bottom culture tubes ( e . g . VWR # 60818 - 667 ) at room temperature .
Place SOC recovery medium in a 37°C water bath .
Pre - warm selective plates at 37°C for 1 hour .
Place electroporation cuvettes ( 1mm ) and microcentrifuge tubes on ice .
As a positive control for transformation , dilute the control pUC19 by 1 : 5 to a final concentration of 10 pg / μl using sterile water .
Thaw NEB Turbo Electrocompetent cells on ice ( about 10 min ) and mix cells by flicking gently .
Transfer 25 μl of the cells ( or the amount specified for the cuvettes ) to a chilled microcentrifuge tube .
Add 1 μl of the DNA solution .
Carefully transfer the cell / DNA mix into a chilled cuvette without introducing bubbles and make sure that the cells deposit across the bottom of the cuvette .
Electroporate using the following conditions for BTX ECM 630 and Bio - Rad GenePulser electroporators : 2.1 kV , 100 Ω , and 25 μF .
The typical time constant is ~ 2.6 milliseconds .
Immediately add 975 µl of 37°C SOC to the cuvette .
Gently mix up and down twice .
Transfer to the 17mm x 100mm round - bottom culture tube .
Shake vigorously ( 250 rpm ) or rotate at 37°C for 1 hour .
Dilute the cells as appropriate then spread 100 - 200 μl cells onto a pre - warmed selective plate .
Incubate plates 8 hours to overnight at 37°C .
