Home  Contacts  Order  OEM  Catalog  Support  Search  Alphabetical Index  Numerical Index  Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification  Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents C A T A L O G

Taq DNA Polymerase (recombinant)

#EP0401 #EP0403 Both supplied with: 10X Taq Buffer with KCl 10X Taq Buffer with (NH4)2SO4 25 mM MgCl2	100 u  (5 u/µl) LC, 100 u (1 u/µl) 0.6 ml 0.6 ml  0.6 ml
#EP0402 #EP0404 Both supplied with: 10X Taq Buffer with KCl 10X Taq Buffer with (NH4)2SO4 25 mM MgCl2	500 u (5 u/µl) LC, 500 u (1 u/µl) 2x1.25 ml 2x1.25 ml  2x1.25 ml
#EP0405 Supplied with: 10X Taq Buffer with KCl 10X Taq Buffer with (NH4)2SO4 25 mM MgCl2	5x500 u (5 u/µl) 10x1.25 ml 10x1.25 ml  10x1.25 ml
#EP0406 Supplied with: 10X Taq Buffer with KCl 10X Taq Buffer with (NH4)2SO4 25 mM MgCl2	10x500 u (5 u/µl) 20x1.25 ml 20x1.25 ml  20x1.25 ml
Related Documents (in pdf, ~40-200 KB): Certificate of Analysis: #EP0401, #EP0402, #EP0403, #EP0404, #EP0405, #EP0406 MSDS (English) MSDS (English-USA) MSDS (German) Flyer
see Protocol for PCR with Taq DNA Polymerase

 

Features

• Thermostable - half life at 95°C is more than 40 minutes.
• Generates PCR products with 3'-dA overhangs.
• Supplied with two buffers - 10X Taq Buffer with KCl and 10X Taq Buffer with (NH₄)₂SO₄. The latter allows for PCR at wide range of magnesium concentrations (see picture below).
• Incorporates modified nucleotides (e.g., biotin-, digoxigenin-, fluorescently-labeled nucleotides).

Fermentas Taq DNA Polymerase, flyer in pdf, 209 KB

Description
The Taq DNA Polymerase is a highly thermostable DNA polymerase from the thermophilic bacterium Thermus aquaticus. The enzyme catalyzes template-dependent polymerization of nucleotides into duplex DNA in the 5'=>3' direction. Taq DNA Polymerase has no detectable 3'=>5' exonuclease (proofreading) activity and possesses low 5'=>3' exonuclease activity. Like other DNA polymerases without 3'=>5' exonuclease activity, Taq DNA Polymerase exhibits deoxynucleotidyl transferase activity, which frequently results in the addition of extra adenines at the 3'-end of PCR products.

Source
E.coli cells with a cloned pol gene from Thermus aquaticus.

Molecular Weight
94 kDa monomer.

Applications

• PCR amplification of DNA fragments as long as 5 kb (1), see protocol.
• Generation of PCR product for TA cloning.
• DNA labeling (2-4).
• DNA sequencing (5).

Quality Control
The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests. Functionally tested in PCR.

Concentration
5 u/µl; 1 u/µl, LC

Definition of Activity Unit
One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30 min at 70°C.
Enzyme activity is assayed in the following mixture: 67 mM Tris-HCl (pH 8.8 at 25°C), 6.7 mM MgCl₂, 1 mM 2-mercaptoethanol, 50 mM NaCl, 0.1 mg/ml BSA, 0.75 mM activated calf thymus DNA, 0.2 mM of each dNTP, 0.4 MBq/ml [³H]dTTP.

Storage Buffer
The enzyme is supplied in: 20 mM Tris-HCl (pH 8.0), 1 mM DTT, 0.1 mM EDTA, 100 mM KCl, 0.5% (v/v) Nonidet P40, 0.5% (v/v) Tween 20 and 50% (v/v) glycerol.

10X Taq Buffer with KCl
100 mM Tris-HCl (pH 8.8 at 25°C), 500 mM KCl, 0.8% (v/v) Nonidet P40.

10X Taq Buffer with (NH₄)₂SO₄
750 mM Tris-HCl (pH 8.8 at 25°C), 200 mM (NH₄)₂SO₄, 0.1% (v/v) Tween 20.

Inhibition and Inactivation

• Inhibitors: ionic detergents (deoxycholate, sarkosyl and SDS) at concentrations higher than 0.06, 0.02 and 0.01%, respectively (6).
• Inactivated by phenol/chloroform extraction.

Note

• The error rate of Taq DNA Polymerase in PCR is 2.2x10^-5 errors per nt per cycle, as determined by a modified method described in (7). Accordingly, the accuracy of PCR is 4.5x10⁴. Accuracy is an inverse of the error rate and shows an average number of correct nucleotides incorporated before an error occurs.
• 10X Taq Buffer without Detergent is recommended for microarray experiments.

Tolerance to different MgCl₂ concentrations in PCR using Taq DNA Polymerase (recombinant) and 10X Taq Buffer with (NH₄)₂SO₄.
A 950 bp single copy gene from human genomic DNA was amplified at a wide range of Mg²⁺ concentrations. M - GeneRuler™ 100 bp Plus DNA Ladder
1-7 - PCR products generated at various MgCl₂ concentrations

Related Products

PCR Master Mix (2X) dNTP Mixes dNTP Set Modified Nucleotides (molecular biology grade): Aminoallyl-dUTP  Biotin-11-dUTP Fluorescein-12-dUTP dm6ATP dm4CTP dm5CTP 10X Taq Buffer without Detergent CloneJET™ PCR Cloning Kit InsTAclone™ PCR Cloning Kit First Strand cDNA Synthesis Kit RevertAid™ First Strand cDNA Synthesis Kits M-MuLV Reverse Transcriptase RevertAid™ M-MuLV Reverse Transcriptase RevertAid™ H Minus M-MuLV Reverse Transcriptase FastRuler™ DNA Ladders, ready-to-use O'RangeRuler™ DNA Ladders, ready-to-use GeneRuler™ DNA Ladders T4 DNA Polymerase Genomic DNA Purification Kit DNA Gel Extraction Kit Agarase Water, nuclease-free

References

1. Innis, M.A., et al., PCR Protocols and Applications: A Laboratory Manual, Academic, New York, 1989.

2. Celeda, D., et al., PCR amplification and simultaneous digoxigenin incorporation of long DNA probes for fluorescence in situ hybridization, BioTechniques, 12, 98-102, 1992.
3. Finckh, U., et al., Producing single-stranded DNA probes with the Taq DNA polymerase: a high yield protocol, BioTechniques, 10, 35-39, 1991.
4. Yu, H. et al., Cyanine dye dUTP analogs for enzymatic labeling of DNA probes, Nucleic Acids Res., 22, 3226-3232, 1994.
5. Innis, M.A., et al., DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA, Proc. Natl. Acad. Sci. USA, 85, 9436-9440, 1988.
6. Weyant, R.S., et al., Effect of ionic and nonionic detergents on the Taq polymerase, BioTechniques, 9, 308-309, 1990.
7. Lundberg, K.S., et al., High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus, Gene, 108, 1-6, 1991.

Purchase of this product 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. For complete license disclaimer, see.

Updated kovo 18, 2008 10:10