Genetic applications of an inverse polymerase chain reaction.
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.
Ochman
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Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110.
Gerber
A S
AS
Hartl
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DL
D001483:Base Sequence; D004251:DNA Transposable Elements; D004270:DNA, Circular / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000235:genetics
D004251:DNA Transposable Elements; D004270:DNA, Circular; D009841:Oligonucleotides; D004259:DNA-Directed DNA Polymerase