Thermo Fisher Scientific
One Shot™ TOP10 Chemically Competent E. coli
C404003
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The target gene was amplified using PCR and after restriction digestion, PCR product was inserted into the pCMV mammalian expression vector. The vector with its load consists of 8kb were transformed into chemically competent E. coliI and incubated in SOC media. Further, the bacterial cells were spreader into agar plate containing selection antibiotics and overnight incubated at 37 degree Celsius. Next day, the bacterial colonies were expanded in selection broth medium and plasmid DNA was tested for the presence of positive DNA clone.
Bacterial transformation
Plasmid DNA, Restriction endonuclease, Incubator, Centrifuge
None
Even at a high plasmid DNA load of 8kb, many colonies were found positive for plasmid DNA with its load. It has a high capacity to uptake bigger DNA and make clones.
Chemically competent E. coli shows a high rate of transformation efficiency
Nothing
Excellent kit for gene cloning purpose