MasterAmp™ DNA Polymerases

MasterAmp™ AmpliTherm™, Taq, Tfl, and Tth DNA Polymerases* are recombinant thermostable DNA polymerases for use in PCR. These enzymes have optimal DNA synthetic activity at temperatures above 70°C and can be used at temperatures up to 95°C. MasterAmp DNA Polymerases are provided with a 10X solution of MasterAmp™ PCR Enhancer with betaine,** which increases the probability of obtaining the desired amplification product, the reproducibility of PCR, and improves the consistency of PCR product yields in multiplex PCR.

Unit Definition: One unit of MasterAmp 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 DNA Polymerases are tested for the absence of detectable nonspecific exo- and endonuclease and RNase activities, and are functionally tested in PCR.

MasterAmp™ AmpliTherm™ DNA Polymerase

Applications

• PCR and multiplex PCR amplification of DNA templates.

MasterAmp™ AmpliTherm™ DNA Polymerase is a proprietary recombinant thermostable DNA polymerase for use in PCR. It lacks both the 5´→3´ structure-dependent exonuclease activity, like that found in Taq DNA polymerase, and the 3´→5´ proofreading exonuclease activity found in some other DNA polymerases. The enzyme is purified to homogeneity using a proprietary procedure that virtually eliminates contaminating bacterial DNA.

MasterAmp™ Taq DNA Polymerase

Applications

• PCR and multiplex PCR amplification of DNA templates.

MasterAmp™ Taq DNA Polymerase is a thermostable DNA polymerase derived from Thermus aquaticus. Having optimal activity at temperatures above 70°C, it was the first thermostable enzyme used for PCR. It has an intrinsic 5´→3´ structure-dependent exonuclease activity, but lacks 3´→5´ proofreading exonuclease activity. The enzyme is highly purified using a proprietary procedure that virtually eliminates contaminating bacterial DNA.

MasterAmp™ Tfl DNA Polymerase

Applications

• PCR and multiplex PCR amplification of DNA templates.
• Long PCR up to 15 kb.
• High-throughput PCR.

Derived from the thermophilic bacterium Thermus flavus, MasterAmp™ Tfl DNA Polymerase is a recombinant DNA polymerase with good thermostability (to ~95°C) and processivity (15-kb PCR products). It is highly purified using a proprietary procedure that virtually eliminates contaminating bacterial DNA.

MasterAmp Tfl 20X PCR Buffer: 1.0 M Tris-HCl (pH 9.0) and 0.4 M ammonium sulfate. A 25 mM solution of MgCl2 is also provided for optimization of the magnesium ion concentration in the reaction. dNTPs are not included; each dNTP should be added to a final concentration of 200 µM for most applications.

MasterAmp™ Tth DNA Polymerase

Applications

• 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.

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.

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.

*Purchase of MasterAmp™ DNA Polymerases includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser’s own internal research. No other patent rights (such as 5´ Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. Covered by issued and pending patents. For complete license statement, see p. iv.

**Covered by issued and/or pending patents.

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