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A Novel PCR-RFLP Detection Method Using an Optimized Set of Restriction Enzymes

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Part of the Neuromethods book series (NM,volume 34)

Abstract

A number of new and useful mutation detection methods have evolved in recent years enabled by the advent of polymerase chain reaction (PCR) (Grompe, 1993). These methods can be divided into two groups: (1) the PCR-based techniques aimed at scanning DNA sequences for unknown mutations and (2) the techniques for identification of known polymorphisms. The essence of the first group is sensitivity, and this group is represented by single-strand conformation polymorphism (SSCP) analysis, denaturing gradient gel electrophoresis (DGGE), heteroduplex analysis (HA), RNase A cleavage, chemical mismatch cleavage, Escherichia coli mismatch repair enzyme-based analysis (Grompe, 1993), and restriction endonuclease fingerprinting (Liu and Sommer, 1995). Once a polymorphism is detected by the techniques described above, there is a clear necessity to switch to a simpler technique. The methods designed for simplicity and speed of genotyping large numbers of individuals are represented by oligodeoxynucleotide hybridization assay, the oligonucleotide ligation assay, allele-specific PCR amplification or, if the DNA variation is detected by a restriction enzyme (RE), PCR-restriction fragment length polymorphism (RFLP) (Grompe, 1993).

Keywords

  • Polymerase Chain Reaction
  • Polymerase Chain Reaction Product
  • Mutation Detection Method
  • Oligonucleotide Ligation Assay
  • Potential Restriction Site

These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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© 1999 Humana Press Inc

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Petronis, A., Timinskas, A., Basile, V., Vicente, A., Janulaitis, A., Kennedy, J.L. (1999). A Novel PCR-RFLP Detection Method Using an Optimized Set of Restriction Enzymes. In: Boulton, A.A., Baker, G.B., Bateson, A.N. (eds) In Vitro Neurochemical Techniques. Neuromethods, vol 34. Humana Press. https://doi.org/10.1385/0-89603-509-3:153

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  • DOI: https://doi.org/10.1385/0-89603-509-3:153

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-509-6

  • Online ISBN: 978-1-59259-639-3

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