 | | | | Taq DNA Polymerase with ThermoPol II (Mg-free) 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 (1,2,3) and a 5´ flap endonuclease activity (4,5).
It is supplied with 10X ThermoPol II (Mg-free) Reaction Buffer and MgSO4. 10X ThermoPol II (Mg-free) Reaction Buffer 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:- PCR
- Primer extension
- Colony PCR
- Long PCR (>5 kb)
Reagents Supplied:
ThermoPol II (Mg-free) 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 kDa
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.
Reagents Supplied with Enzyme:
10X ThermoPol II (Mg-free) Reaction Buffer
100 mM MgSO4
Reaction Conditions:
1X ThermoPol II (Mg-free) Reaction Buffer, DNA template, primers, 200 µM dNTPs (not included), 2 mM MgSO4 and 1.25 units of Taq DNA Polymerase in a total reaction volume of 50 µl.
1X ThermoPol II (Mg-free) Reaction Buffer:
20 mM Tris-HCl
10 mM (NH4)2S04
10 mM KCl
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
- Does the presence of Ca2+ inhibit PCR reactions?
Protocols
Protocols for Taq DNA Polymerase with ThermoPol II (Mg-free) 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 II (Mg-free) 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, 0.2 µM primers and 2 mM MgSO4 in ThermoPol II (Mg-free) Reaction Buffer results in the expected 5 kb 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.
References
- 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 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 II (Mg-free) 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 Reaction Buffer Pack
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