 | | | | Taq DNA Polymerase with ThermoPol Buffer | | | |
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Prices are in US dollars and valid only for US orders.
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- Isolated from a recombinant source
- Supplied with 10X Reaction Buffer
- 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:
Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity and a double-strand specific 5´→3´ exonuclease activity.
It is supplied with 10X ThermoPol Reaction Buffer, which contains a nonionic detergent to increase enzyme stability during longer incubations.
Source:
An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1
Applications:- Primer extension
- PCR
- Colony PCR
Reagents Supplied:
ThermoPol Reaction Buffer Pack (10X)
Enzyme Properties
Polymerase Properties | Thermophilic Polymerase Characteristics
3´ to 5´ Exonuclease: No
5´ to 3´ Exonuclease: Yes
Strand Displacement: No
Error Rate: 285
x10-6 bases
Heat Inactivation:
No
Molecular Weight:
Theoretical: 94,000 daltons
Reaction & Storage Conditions
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 15 nmol of dNTP into acid-insoluble material in 30 minutes at 75°C.
Unit Assay Conditions:
1X ThermoPol Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 200 µg/ml activated Calf Thymus DNA.
Reaction Conditions: 1X ThermoPol Reaction Buffer, DNA template, primers, 200 µM dNTPs (not included) and 1.25 units of Taq DNA Polymerase in a total reaction volume of 50 µl.
1X ThermoPol Reaction Buffer:
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton X-100
pH 8.8 @ 25°C
Concentration:
5,000 units/ml
Storage Conditions:
10 mM Tris-HCl
100 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
0.5% Tween-20
0.5% NP-40
pH 7.4 @ 25°C
Storage Temperature:
-20°C
FAQs
- Can Taq DNA Polymerase be used in other buffers?
- Why is the product a smear when visualized on an agarose gel?
- Why is there no product when visualized on an agarose gel?
- How should I set up a PCR reaction using Taq DNA Polymerase?
- The product sequence doesn't completely match the expected sequence. How can this result be improved?
- What type of DNA ends result from a primer extension reaction or a PCR reaction using LongAmp Taq DNA Polymerase?
- When should Taq DNA Polymerase be used in a primer extension reaction or for PCR?
- Will the 5'→3' exonuclease activity of Taq DNA Polymerase degrade primers?
- Can Taq DNA Polymerase be used for nick translation?
- Does the presence of Ca2+ inhibit PCR reactions?
Protocols
Protocols for Taq DNA Polymerase with ThermoPol Buffer
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Endonuclease Activity:
Incubation of a 50 μl reaction containing 20 units of Taq DNA Polymerase with ThermoPol Buffer with 1 μg of
ΦX174 DNA for 4 hours at 75ºC resulted
in < 10% conversion to RFII as determined by agarose gel electrophoresis.
5 kb Lambda PCR:
25 cycles of PCR amplification of 5 ng Lambda DNA with 1.25 units of Taq DNA Polymerase in the presence of 200 µM dNTPS and 0.2 µM primers in ThermoPol Reaction Buffer results in the expected 5 kb product.
3´→ 5´ Exonuclease Activity:
Incubation of 20 units of Taq DNA Polymerase with ThermoPol Buffer in 20 µl of a 10 nM solution of a fluorescent 5’-FAM labeled oligonucleotide for 30 minutes at 37°C or 75°C yields no detectable 3’-->5’ degradation as determined by capillary electrophoresis.
References
- Chien, A., Edgar, D.B. and Trela, J.M. (1976) J. Bact., 127, 1550-1557.
- Kaledin, A.S., Sliusarenko, A.G. and Gorodetskii, S.I. (1980) Biokhimiya, 45, 644-651.
- Lawyer, F.C. et al. (1993) PCR Methods 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.
- 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.
Reagents Sold Separately
ThermoPol Reaction Buffer Pack
Companion Products
Taq 5X Master Mix
Taq 2X Master Mix
Taq PCR Kit
Deoxynucleotide Solution Mix
Deoxynucleotide Solution Set
Diluent F
Magnesium Sulfate (MgSO4) Solution
Quick-Load® Taq 2X Master Mix
ThermoPol DF (Detergent-free) Reaction Buffer Pack
ThermoPol II (Mg-free) Reaction Buffer Pack
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