Genetics 1988 Nov;120(3):621-3

Genetic applications of an inverse polymerase chain reaction.

Ochman H, Gerber AS, Hartl DL

Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110.

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.

MeSH Terms:

• Base Sequence
• DNA Transposable Elements
• DNA-Directed DNA Polymerase/genetics*
• DNA, Circular/genetics
• Escherichia coli/genetics*
• Gene Amplification/methods*
• Molecular Sequence Data
• Oligonucleotides/genetics
• Oligonucleotides/chemical synthesis

Substances:

• Oligonucleotides
• DNA, Circular
• DNA Transposable Elements
• DNA-Directed DNA Polymerase

PMID: 2852134, UI: 89137892