LongAmp Hot Start Taq DNA Polymerase is a unique blend of aptamer-based Hot Start Taq and Deep VentR™ DNA Polymerases. The aptamer-based inhibitor binds reversibly to the enzyme, inhibiting polymerase activity at temperatures below 45°C, but releases the enzyme during normal PCR cycling conditions. This permits assembly of PCR reactions at room temperature. An advantage of the aptamer-based hot start mechanism is that it does not require a separate high temperature incubation step to activate the enzyme. The 3´→5´ exonuclease activity of Deep VentR DNA Polymerase increases the fidelity and robust amplification of Hot Start Taq DNA Polymerase (1). LongAmp Hot Start Taq DNA Polymerase offers two-fold higher fidelity than Hot Start Taq DNA Polymerase alone. A wide range of PCR products can be generated; up to 30 kb from lambda or human genomic DNA.
- Room temperature reaction setup
- More robust & longer amplicons than Taq
Product SourceAn E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1 and an E. coli strain that carries the Deep VentR DNA Polymerase gene from Pyrococcus species GB-D.
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|LongAmp™ Taq Reaction Buffer Pack||-20||5X|
Advantages and Features
- High-specificity Long Range PCR
- Colony PCR
Properties and Usage
Unit DefinitionOne unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 75°C.
1X LongAmp Taq Reaction Buffer
1X LongAmp™ Taq Reaction Buffer:
60 mM Tris-SO4
20 mM (NH4)2SO4
0.06% IGEPAL® CA-630
0.05% Tween® 20
2 mM MgSO4
pH 9 @ 25°C
10 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
0.5% Tween® 20
0.5% IGEPAL® CA-630
pH 7.4 @ 25°C
5' - 3' ExonucleaseYes
3' - 5' ExonucleaseYes
Unit Assay Conditions1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA.
Quality Control AssaysThe following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
- Inhibition of Primer Extension (Hot Start, Radioactivity Incorporation):
The hot start polymerase is compared to the polymerase in non-hot start conditions using primer extension; inhibition is assessed by comparing radioactive incorporation.
- PCR Amplification (DNA Polymerase):
The polymerase is tested in a polymerase chain reaction (PCR) using a control template and specific primers, resulting in the expected product.
- PCR Amplification (Hot Start, Human Genomic DNA):
The polymerase is tested in a hot start polymerase chain reaction (PCR) using Human genomic DNA as the control template and specific primers, resulting in an increase in yield of the expected product and a decrease in non-specific genomic bands when compared to a non-hot start control reaction.
LicensesNotice to Purchaser:
Nucleic acid-based aptamers for use with thermophilic DNA polymerases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. (See Patent Nos. 5,475,096; 5,670,637; 5,696,249 5,874,557; and 5,693,502). New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the aptamer-based LongAmp Hot Start Taq DNA Polymerase for RESEARCH PURPOSES ONLY. Commercial use of the aptamer-based LongAmp Hot Start Taq DNA Polymerase requires a license from New England Biolabs, Inc. Please contact firstname.lastname@example.org for more information. Patent Nos. 5,475,096; 5,670,637; 5,696,249 5,874,557; and 5,693,502
TrademarksLONGAMP® and THERMOPOL® are registered trademarks of New England Biolabs, Inc.
DEEP VENT™ is a trademark of New England Biolabs, Inc.
IGEPAL® is a registered trademark of Rhodia Operations.
TWEEN® is registered trademark of Uniqema Americas LLC.
- 5'→3' flap endonuclease destroys displaced strand.
- Barnes, W.M. (1994). Proc. Natl. Acad. Sci. USA. 91, 2216-2220.
- Saiki R.K. et al. (1985). Science. 230, 1350-1354.
- Powell, L.M. et al. (1987). Cell. 50, 831-840.
- Sun, Y., Hegamyer, G. and Colburn, N. (1993). Biotechniques. 15, 372-374.
- Sarkar, G., Kapelner, S. and Sommer, S.S. (1990). Nucleic Acids Res.. 18, 7465.
- What is the recommended enzyme amount when using LongAmp™ Hot Start Taq DNA Polymerase?
- What is the fidelity of the LongAmp™ Hot Start Taq DNA Polymerase compared to Taq DNA Polymerase?
- Can the extension step be carried out at 72°C when using LongAmp™ Hot Start Taq DNA Polymerase?
- What type of DNA ends result from a primer extension reaction or a PCR using LongAmp™ Hot Start Taq DNA Polymerase?
- What is the extension rate when using LongAmp™ Hot Start Taq DNA Polymerase?
- Why is the product a smear when visualized on an agarose gel?
- Can LongAmp™ Hot Start Taq DNA Polymerase be used to amplify GC-rich amplicons?