Tth DNA Polymerase is a thermostable enzyme of approximately 94,000
daltons isolated from Thermus thermophilus HB-8. Tth DNA Polymerass catalyzes the polymerization of nucleotides
into duplex DNA in the 5' 3' direction in the presence of MgCl2. The enzyme is also capable of catalyzing the polymerization
of DNA using an RNA template in the presence of MnCl2. The ability of Tth DNA Polymerass to reverse transcribe at elevated
temperatures minimizes the problems encountered with strong secondary
structures in RNA since they are unstable at higher reaction temperatures.
Higher temperatures also result in increased specificity of primer
hybridization and extension.
10 X Reverse Transcription Buffer
100 mM Tris-HCl (pH 8.3) 900 mM KCl
10 X Polymerase Reaction Buffer
500 mM KCl 100 mM Tris-HCl (pH 9.0) 1 % Tritons X-100
Buffer is optimized for use with 0.2 mM for each of the dNTPs.
REFERENCES
Rottimann, C et al. (1985) Eur. J. Biochem. 149: 41.