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.

Applications include:
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.