During “cut and paste” cloning, the PCR-amplified gene is cut by restriction enzymes. How does the...

During “cut and paste” cloning, the PCR-amplified gene is cut by restriction enzymes. How does the synthesis of DNA oligonucleotide primers facilitate the ability to ‘cut’ the amplified gene?

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the gene which is to be inserted into the plasmid may not have the required restriction enzymes sites at the ends, so primers can be designed such that it has the restriction site, primer binds to the template strand and DNA polymerase adds new nucleotides to the 3`-OH of the primer, so primer remains in the amplified DNA, after many rounds of replication the amplified DNA will have the restriction sites which is added in the primer at the ends, so the amplified DNA can be digested with those enzymes and the plasmid is also digested with same restriction enzymes and then the amplified DNA and the plasmid can be ligated using DNA ligase.

gladiator answered 1 year ago

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