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Taq 5X Master Mix |
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Prices are in US dollars and valid only for US orders.
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Description:
Taq 5X Master Mix is an optimized ready-to-use solution containing Taq DNA Polymerase, dNTPs, MgCl2, KCl and stabilizers. It is best for routine PCR applications from templates including pure DNA solutions, bacterial colonies, and cDNA. It can amplify up to 4 kb from complex genomic DNA or up to 5 kb from lambda DNA.
Taq DNA Polymerase is a thermostable DNA polymerase that posesses a 5´→ 3´ polymerase activity and a double-strand specific 5´→ 3´ exonuclease activity.
Source:
An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1.
Applications:- PCR
- Primer Extension
- Microarray Analysis
- High-Throughput PCR
Reagents Supplied:
MgCl2 (25 mM)
Taq MasterMix
Enzyme Properties
Heat Inactivation:
No
Reaction & Storage Conditions
Reaction Conditions:
1X Taq MasterMix
Incubate at
75°C.
1X Taq MasterMix:
10 mM Tris-HCl
50 mM KCl
1.5 mM MgCl2
0.2 mM dNTPs
5 % Glycerol
0.08 % NP-40
0.05 % Tween-20
25 units/ml Taq DNA Polymerase
pH 8.6 @ 25°C
Unit Definition:
One unit of Taq DNA Polymerase is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 75°C.
Unit Assay Conditions:
1X Standard Taq Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 200 µg/ml activated Calf Thymus DNA.
Concentration:
5 X
Storage Temperature:
-20°C
Notes
General notes:- Keep at -20°C for long term storage. Taq 5X Master Mix is stable at 4°C for six months or for fifteen freeze-thaw cycles. For daily use, we recommend keeping an aliquot at 4°C.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
3' to 5' Exonuclease Activity:
No detectable 3' → 5' nuclease activity was observed when 20 units
of Taq 5X Master Mix was incubated with substrates containing either 3' extensions or blunt ends.
Endonuclease Activity:
Incubation of a 50 μl reaction containing 20 units of Taq 5X Master Mix with 1 μg of
ΦX174 RF I 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 2.5 units of Taq DNA Polymerase in the presence of 200 µM dNTPS, 0.2 µM primers and 1X Standard Taq Reaction Buffer resulted in a yield of 100 ng specific product.
References
- Henke et al. (1997) Nucleic Acids Res., 25, 3957-3958.
- Sun et al. (1993) Biotechniques, 15, 372-374.
- Sarkar et al. (1990) Nucleic Acids Res., 18, 7465.
- Don, R.H. et al. (1991) Nucleic Acids Res., 19, 4008.
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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