Collect recombinant DNA, uncut plasmid as control, SOC medium, and competent bacterial cells. Thaw the frozen competent cells and transfer 100 microliters to a chilled tube.
▶ 00:02
2
Add DNA to competent cells
Add recombinant DNA to the first tube of competent cells. Prepare a control tube with the same volume of cells and add uncut plasmid instead.
▶ 00:40
3
Incubate cells on ice
Immediately place both tubes on ice and incubate for 10 minutes to allow DNA uptake.
▶ 00:55
4
Heat shock the cells
Transfer tubes to a 42 degrees Celsius water bath and heat shock for 45 to 50 seconds to facilitate DNA entry into cells.
▶ 01:02
5
Cool cells on ice again
Place the tubes back on ice for another 2 minutes to allow cells to recover.
▶ 01:09
6
Add SOC medium and recover
Add 900 microliters of cold SOC medium to each tube and incubate at 37 degrees Celsius with shaking for 60 minutes to allow cells to express antibiotic resistance genes.
▶ 01:14
7
Prepare transformed cells for plating
The transformed cells are now ready for dilution and plating on selective media to identify successfully transformed colonies.
▶ 01:25
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