Genetic applications of an inverse polymerase chain reaction.

Howard Ochman, Anne S. Gerber, Daniel L. Hartl

Genetics · 1988 · 1.9K citations · 10 references

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TL;DR

Inverse PCR (IPCR) is a versatile technique in molecular genetics for amplifying and identifying sequences flanking transposable elements. The study presents a rapid in vitro method to amplify DNA sequences flanking a known region, demonstrating IPCR’s feasibility by targeting an IS1 element in *E. coli*. The method employs PCR with reverse‑oriented primers on a circularized restriction fragment.

Abstract

Abstract A method is presented for the rapid in vitro amplification of DNA sequences that flank a region of known sequence. The method uses the polymerase chain reaction (PCR), but it has the primers oriented in the reverse direction of the usual orientation. The template for the reverse primers is a restriction fragment that has been ligated upon itself to form a circle. This procedure of inverse PCR (IPCR) has many applications in molecular genetics, for example, the amplification and identification of sequences flanking transposable elements. In this paper we show the feasibility of IPCR by amplifying the sequences that flank an IS1 element in the genome of a natural isolate of Escherichia coli.

References

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