MasterAmp™ Tfl DNA Polymerase
25mM MgCl2
| Catalogue number | Pack size |
MasterAmp™ 10X PCR Enhancer
| Catalogue number | Pack size |
MasterAmp™ Tfl DNA Polymerase
| Catalogue number | Pack size |
MasterAmp™ Tfl DNA Polymerase - Enzyme Only
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Covered by issued and pending patents.
* Epicentre's® PCR products are sold under licensing arrangements with F. Hoffmann-La Roche Ltd., Roche Molecular Systems, Inc., and Applied Biosystems.
The products containing a thermostable DNA polymerase are accompanied by a limited license to use it in the Polymerase Chain Reaction (PCR) and RT-PCR for life science research in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by the up-front license fee, either by payment to Perkin-Elmer or as purchased, i.e., an authorised thermal cycler.
Last modified: 2008-07-15 14:57:15
Includes MasterAmp™ PCR Enhancer (with betaine) for improved PCR
Derived from the thermophilic bacterium Thermus flavus, MasterAmp™ Tfl DNA Polymerase is a thermostable DNA polymerase useful for performing PCR. It has optimal enzymatic activity at temperatures above 70°C, and can be used at temperatures up to 95™C. Epicentre®'s MasterAmp™ Tfl DNA Polymerase has been shown to produce large PCR products (15kb), and is highly purified using a proprietary procedure that virtually eliminates contaminating bacterial DNA. A 10X solution of MasterAmp™ PCR Enhancer (with betaine) is provided with the enzyme. The MasterAmp PCR Enhancement Technology* substantially increases both the probability of obtaining the desired amplification product and the reaction-to-reaction consistency of PCR. It also greatly improves the consistency of PCR product yields in multiplex PCR.
Unit Definition:
One unit of MasterAmp™ Tfl DNA Polymerase converts 10nmoles of dNTPs into acid-insoluble material in 30 minutes at 70°C under standard assay conditions.
Storage Buffer:
50% glycerol containing 50mM Tris-HCl (pH 7.5), 100mM NaCl, 0.1mM EDTA, 0.5% Tween 20, 0.5% NP-40, and 1mM DTT.
MasterAmp™ Tfl 20X PCR Buffer:
1.0M Tris-HCl (pH 9.0) and 0.4M ammonium sulphate. A 25mM solution of MgCl2 is also provided for optimisation of the magnesium ion concentration in the reaction. dNTPs are not included; each dNTP should be added to a final concentration of 200µM for most applications.
Optimisation of [Mg2+]:
Mg2+ is required for activity of MasterAmp Tfl DNA Polymerase. We recommend that initial experiments be performed at 1.5 mM MgCl2. The Mg2+ concentration may have to be adjusted for some template/primer pair combinations, or for dNTP concentrations other than 200µM each. Alternatively, we recommend the MasterAmp PCR Optimisation Kit with ammonium sulphate.
Quality Control:
MasterAmp™ Tfl DNA Polymerase is tested for the absence of detectable nonspecific DNA exo- and endonuclease and RNase activities, and is functionally tested in PCR.
Last modified: 2008-05-14 16:46:46
Applications
- PCR amplification of DNA templates
- Multiplex PCR
Benefits
- Improved amplification of difficult-to-amplify templates
- Produces PCR products up to 15kb
- Adaptable to high-throughput PCR formats1
Last modified: 2008-05-14 16:46:46
Protocols for: MasterAmp™ Tfl DNA Polymerase
MasterAmp™ Tfl DNA Polymerase Protocol
(catalogue number F72250 / F72500 / F7205K / F72250N / FB3715 / FB3760)
MasterAmp™ 10X PCR Enhancer Protocol
(catalogue number ME81201 / ME81205 / ME81210)
Please note: all protocols off site are the responsibility of the products supplier
Last modified: 2008-05-14 16:46:46
References
- Grujic, D. et al. (1994) BioTechniques 17, 291.
Last modified: 2008-05-14 16:46:46
