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Prices are in US dollars and valid only for US orders.
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- Robust and reliable reactions
- Tolerates a wide range of templates
- Incorporates dUTP, dITP and fluorescently-labeled nucleotides
- Exceptional value in terms of cost per unit
Description:
The Polymerase Chain Reaction (PCR) is a powerful and sensitive technique for DNA amplification (1). PCR amplifies specific DNA sequences exponentially by using multiple cycles of a three-step process. First, the double-stranded DNA template is denatured at a high temperature. Sequence-specific primers are then annealed to sites flanking the target sequence. A thermostable DNA polymerase, such as Taq DNA Polymerase (2–6), then extends the annealed primers, thereby doubling the amount of the original DNA sequence. This newly synthesized product then becomes an additional template for subsequent cycles of amplification. These three steps are repeated for 20 to 30 cycles, resulting in a 105 –109 fold increase in target DNA concentration.
Kit Components:
Taq DNA Polymerase with Standard Taq Buffer (5,000 units/ml)
Deoxynucleotide Solution Mix (200 μl)
MgCl2 (25 mM)
Quick-Load® 2-Log DNA Ladder(0.1-10.0 kb)
Standard Taq (Mg-free) Reaction Buffer Pack (10X)
Storage Conditions
Storage Temperature:
-20°C
Protocols
Protocols for Taq PCR Kit
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
3´→5´ Exonuclease Activity:
Incubation of a 20 μl reaction in ThermoPol Reaction Buffer containing a minimum of 20 units of Taq DNA Polymerase with 10 nM fluorescent internally labeled oligonucleotide for 30 minutes at either 37°C or 75°C yields no detectable 3´→5´ degradation as determined by capillary electrophoresis.
Endonuclease Activity:
Incubation of a 50 μl reaction in ThermoPol Reaction Buffer containing a minimum of 20 units of Taq DNA Polymerase with 1 μg of supercoiled ΦX174 RF I DNA for 4 hours at 75°C results in < 10% conversion to the nicked form as determined by agarose gel electrophoresis.
Reaction Buffers
The supplied NEB reaction buffers and supplements are free of detectable nucleases.
Deoxynucleotide Solution
Deoxynucleotide solutions are certified free of nucleases and phosphatases.
5´ kb Lambda PCR:
25 cycles of PCR amplification of 5 ng Lambda DNA with 2.5 units of Taq DNA Polymerase in the presence of 200 μM dNTPs and 0.2 μM primers in Standard Taq Reaction Buffer results in the expected 5 kb product.
PCR:
Taq PCR Kits are qualified for use in PCR by demonstrating the ability to amplify both a 500 bp single-copy human gene and an 8 kb Lambda amplicon in a 25 cycle reaction.
References
- Saiki, R.K. et al. (1985) Science, 230, 1350-1354.
- Chien, A., Edgar, D.B. and Trela, J.M. (1976) J. Bact., 127, 1550-1557.
- Kaledin, A.S., Slyusarenko, A.G. and Gorodetskii, S.I. (1980) Biokhimiya, 45, 644-651.
- Lawyer, F.C. et al. (1993) PCR Method and Appl., 2, 275-287.
- Longley, M.J., Bennett, S.E. and Mosbaugh D.W. (1990) Nucleic Acids Res., 18, 7317-7322.
- Lyamichev, V., Brow, M.A. and Dahlberg, J.E. (1993) Science, 260, 778-783.
- Powell, L.M. et al. (1987) Cell, 50, 831-840.
Reagents Sold Separately
Taq DNA Polymerase with Standard Taq Buffer
Deoxynucleotide Solution Mix
Quick-Load® 2-Log DNA Ladder(0.1-10.0 kb)
Standard Taq (Mg-free) Reaction Buffer Pack
Companion Products
Taq DNA Polymerase with Standard Taq (Mg-free) Buffer
Taq DNA Polymerase with ThermoPol II (Mg-free) Buffer
Taq 2X Master Mix
Taq DNA Polymerase with ThermoPol Buffer
Taq PCR Kit with Controls
Deoxynucleotide Solution Set
Diluent F
DyNAzyme™ II Hot Start DNA Polymerase
Phire® Hot Start DNA Polymerase
Phusion® Hot Start II High-Fidelity DNA Polymerase
Quick-Load® Taq 2X Master Mix
ThermoPol DF (Detergent-free) Reaction Buffer
ThermoPol II (Mg-free) Reaction Buffer Pack
ThermoPol Reaction Buffer
Legal
Licenses/Patents/Disclaimers:
Some applications in which this product can be used may be covered by patents issued and applicable in the United States and certain other countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a patent license depending upon the particular application in which the product is used.
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