TRANSFORMATION protocol of DH5α Competent E. coli cells

1-5 μl to the cells (1 to 10 ng DNA) is added to the cell and mixed (flicking the tube /not with pipette)

Incubate the mixture in ice for 30 minutes

Heat shock for 20 seconds in 42°C water bath

Place tubes in ice immediately for 2 minutes.

Add 400 µl of LB media / SOC in the tube.

Incubate the tubes at 37°C shaker for 1 hour at 225 rpm.

After incubation spread 100 µl on the LB (Ampicillin) plates or other selective agar plates.

Incubate plates at 37°C overnight in incubator.

Binod G C

I'm Binod G C (MSc), a PhD candidate in cell and molecular biology who works as a biology educator and enjoys scientific blogging. My proclivity for blogging is intended to make notes and study materials more accessible to students.

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