A distinctive property of Tth DNA polymerase: enzymatic amplification in the presence of phenol.
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.
Katcher
H L
HL
Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla 10595.
Schwartz
I
I
D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000302:isolation & purification; D004259:DNA-Directed DNA Polymerase / Q000494:pharmacology*; D019800:Phenol; D010636:Phenols; D016133:Polymerase Chain Reaction; D012335:RNA, Ribosomal / Q000235:genetics
D010636:Phenols; D012335:RNA, Ribosomal; D019800:Phenol; D004259:DNA-Directed DNA Polymerase