Question

You want to check if a plasmid in E. coli contains a specific gene of interest. If you are given a liquid culture of this strain, what should you do?

Plasmid isolation, PCR, gel electrophoresis
Plasmid isolation, PCR, transformation
Plasmid isolation, ligation, PCR, gel electrophoresis
Plasmid isolation, transformation into E. coli DH5a, digestion with BamHI

Answer 1

Plasmid isolation, PCR and gel elctrophoresis

Plasmid is isolated by cell lysis using SDS or NaOH. Then precipitation of proteins and chromosomal DNA is done using rapid acidification. After plasmid isolation, specific primers are designed according to the gene of interest. After primer designing PCR (polymerase chain reaction) should be done. Then PCR product should run on the agrose gel. Band will appear on the gel if the specific gene of interest is present on the plasmid of e.coli.

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