Inverse Polymerase Chain Reaction

  • Daniel L. Hartl
  • Howard Ochman
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 58)

Abstract

The inverse polymerase chain reaction (IPCR) was the first extension of the conventional polymerase chain reaction to allow the amplification of unknown nucleotide sequences without recourse to conventional cloning. In the conventional polymerase chain reaction (PCR), synthetic oligonucleotides complementary to the ends of a known sequence are used to amplify the sequence (1,2). The primers are oriented with their 3′ ends facing each other, and the elongation of one primer creates a template for annealing the other primer. Repeated rounds of primer annealing, polymerization, and denaturation result in a geometric increase in the number of copies of the target sequence. However, regions outside the boundaries of the known sequence are inaccessible to direct amplification by PCR. Since DNA synthesis oriented toward a flanking region is not complemented by synthesis from the other direction, there is at most a linear increase in the number of copies of the flanking sequence.

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Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Daniel L. Hartl
    • 1
  • Howard Ochman
    • 2
  1. 1.Department of Organismic and Evolutionary BiologyHarvard UniversityCambridgeUK
  2. 2.Department of GeneticsWashington University School of MedicineSt. Louis

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