A distinctive property of Tth DNA polymerase: enzymatic amplification in the presence of phenol

Biotechniques. 1994 Jan;16(1):84-92.

Abstract

The ability of thermostable DNA polymerases to mediate template-dependent DNA synthesis in the presence of phenol has been examined as monitored by amplification of a specific Borrelia burgdorferi rRNA sequence. Tth DNA polymerase displayed the unique property of maintaining both DNA- and RNA-dependent DNA polymerase activities in the presence of 2%-5% (vol/vol) of phenol-saturated PBS buffer. Tth DNA polymerase mediated reverse transcriptase activity was unaffected by phenol-saturated phosphate-buffered saline concentrations as high as 15% (vol/vol). By contrast, Taq DNA Polymerase was inactive under these conditions. The ability to function in the presence of phenol can greatly simplify reverse transcriptase, PCR and reverse transcription-PCR protocols since the phenol-saturated aqueous phase of a phenol partition can be added directly to the reaction mixtures. The simplicity of the procedures described should have applicability to a broad range of basic research, clinical and forensic applications.

Publication types

  • Research Support, Non-U.S. Gov't
  • Technical Report

MeSH terms

  • Borrelia burgdorferi Group / genetics
  • Borrelia burgdorferi Group / isolation & purification
  • DNA-Directed DNA Polymerase / pharmacology*
  • Phenol
  • Phenols*
  • Polymerase Chain Reaction*
  • RNA, Ribosomal / genetics

Substances

  • Phenols
  • RNA, Ribosomal
  • Phenol
  • DNA-Directed DNA Polymerase