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.