Tth 111 Reverse Transcriptase Thermus thermopilus Reverse Transcriptase catalyzes the reverse transcription of RNA to cDNA at elevated temperatures in the presence of Mn2+.Source: Thermus thermophilusDescription:• Suitable for high temperature synthesis of DNA (1).• Synthesizes cDNA from RNA template (2).• Results in greater specificity of primer hybridization and extension of RNA.• Can reverse transcribe at elevated temperatures (2).• Minimizes problems with strong secondary structure of RNA.• Used for efficient PCR of DNA, containing problematic secondary structures.• Applicable to RT-PCR; the same enzyme is used for both reverse transcription and following amplification of obtained cDNA template (2).• Resistant to amplification inhibitors present in template DNA isolated from problematic samples (3,4). Unit Definition:one unit is the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insoluble form in 10 min at 50°C.Storage Conditions: Store at –20°C.Storage Buffer: 50 mM Tris-HCl (pH 7.5 at 22°C), 5 mM dithiothreitol, 0.1 mM EDTA, 50% (v/v) glycerol and stabilizers.Assay Conditions: 40 mM Tris-HCl (pH 8.5 at 22°C), 1 mM MnCl2, 1 mg/ml bovine serum albumin, 10 mM dithiothreitol, 0.5 mM [α-³²P] dTTP and 0.4 mM poly(A)·(dT)50. Incubation is at 50°C for 10 min in a reaction volume of 50 µl.Quality Control: All preparations are assayed for contaminating endonuclease, exonuclease, nonspecific RNase and single- and double-stranded DNase activities.References:1. Wang, A. M., Doyle, M.V. and Mark, D.F. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 9717-9721.2. Myers, T. W., Gelfanld, D. H. (1991) Biochemistry 30, 7661-7666.3. Kather, H. L., Schwartz, I. (1994) Biotechniques 16, 84-92.4. Poddar, S. K., Sawyer, M. H., Connor, J. D. (1998) J. Med. Microbiol. 47, 1131-1135.