A thermostable DNA- and RNA-dependent (i.e., reverse transcriptase) DNA polymerase for both PCR and RT-PCR applications
- Flexible: Use a single enzyme for both PCR and RT-PCR applications, greatly simplifying reaction optimization and setup
- Thermostable: Enables amplification of DNA and RNA targets containing high degrees of secondary structure
MasterAmp™ Tth DNA Polymerase is a recombinant DNA enzyme from Thermus thermophilus that has both DNA-dependent and RNA-dependent (i.e., reverse transcriptase) DNA polymerase activities up to ~95°C. MasterAmp Tth DNA Polymerase is highly purified using a proprietary procedure that virtually eliminates contaminating bacterial DNA.
- PCR amplification of DNA and one-step RT-PCR of RNA.
- PCR amplification of RNA and DNA templates having a high degree of secondary structure.
Optimization of [Mg2+] and [Mn2+]: Magnesium and manganese ion concentrations should be optimized for DNA synthesis. The optimal concentration of each metal cation is highly dependent on the dNTP concentration and on the template, primers, and protocol used. For many templates, the optimal concentration of magnesium ions using MasterAmp Tth DNA Polymerase is approximately 1.5 mM if the concentration of each dNTP in the reaction is 0.2 mM. If used with 1.5 mM Mg2+, the optimal concentration of Mn2+ for RNA-dependent DNA synthesis is approximately 0.5 mM for many templates. For an RT-PCR reaction, we recommend 3.0 mM Mg2+ and 0.5 mM Mn2+ for most templates. Alternatively, we recommend the MasterAmp™ PCR Optimization Kit with ammonium sulfate for rapid PCR optimization.
Unit Definition: One unit of MasterAmp Tth DNA Polymerase converts 10 nmol of dNTPs into acid-insoluble material in 30 minutes at 70°C under standard assay conditions.
Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 0.5% Tween 20, 0.5% NP-40, and 1 mM DTT.
Quality Control: MasterAmp Tth DNA Polymerase is tested for the absence of detectable nonspecific exo- and endonuclease and RNase activities, and are functionally tested in PCR.