Plasmid transduction using competent cell
An.Huang
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Abstract
Plasmid can be transduced into bacteria at competent state using heat shock. This protocol helps transduce plasmid into competent cells.
Steps
Take competent cell out from -80℃ fridge and thaw on ice.
When the cells are completely thawed, pipette 2µL plasmid DNA solution into 100µL competent cell.
Put the cell in ice for 0h 30m 0s
Conduct heat shock on the competent cell by placing the cell in 42°C water bath for 0h 1m 30s.
Put the cells back into ice for 0h 2m 0s
Add 900µL LB broth medium into competent cell mixture. Shake at 180rpm for 0h 45m 0s
Centrifuge at 6000rpm.
Discard 900µL supernatant and resuspend the pellet in the rest 100µL supernatant.
Spread the cells onto LB agar plates.
Place the plate with lid on upside for 1h 0m 0s.
Invert the plate and culture at 37°C in a biomedical incubator overnight.
