Inverse Polymerase Chain Reaction, Clone, Gene, DNA, Polymerization, Cloning of Foreign DNA, Cloning of Flanking Sequences

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Inverse PCR

In this technique, the amplification of those DNA sequences takes place, which are away from the primers and not those which are flanked by the primers. For instance, if the border sequences of a DNA segment are not known and those of a vector are known, then the sequence to be amplified may be cloned in the vector and border sequences of vector may be used as primers in such a way that the polymerization proceeds in inverse direction i.e. away from the vector sequence flanked by the primers and towards the DNA sequence of inserted segment. Similarly, if the gene sequence is known, one can use its border sequences as primers, for an inverse PCR reaction, to amplify the sequences flanking this gene e.g. the regulatory sequences.

Inverse PCR

Inverse PCR (a) amplification of genes using flanking vector sequences as primers (b) amplification of sequences flanking the gene, using sequences at the two ends of the gene as primers

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