Description:
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Taq PCR Master Mix (2x) is a ready-to-use solution containing Taq DNA Polymerase, optimized reaction buffer, MgCl2 and dNTPs.
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Use of Taq PCR Master Mix (2x) allows to save time and reduce contamination risk due to fewer pipetting steps during PCR set-up.
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Taq DNA Polymerase is a thermostable enzyme of approximately 94 kDa from Thermus aquaticus.
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Ultrapure, recombinant protein.
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The enzyme replicates DNA at 74°C and exhibits a half-life of 40 min at 95°C (1,2).
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Catalyzes the polymerization of nucleotides into duplex DNA in the 5´->3´ direction in the presence of magnesium ions.
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Maintains the 5´->3´ exonuclease activity.
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Lacks the 3´->5´ exonuclease activity.
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Adds extra A at the 3' ends.
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Taq PCR Master Mix (2x) is recommended for use in PCR and primer extension reactions at elevated temperatures to obtain a wide range of DNA products up to 10 kb.
Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmoles of dNTP into acid-insoluble material in 30 min at 74°C. The reaction conditions are: 50 mM Tris-HCl (pH 9.0 at 25°C), 50 mM NaCl, 5 mM MgCl2, 200 μM each of dATP, dCTP, dGTP, dTTP (a mix of unlabeled and [³H]dTTP), 10 μg activated calf thymus DNA and 0.1 mg/ml BSA in a final volume of 50 μl.
Storage Conditions: Store at -20°C for long-term storage or at 4°C for up to 2 months.
Taq PCR Master Mix (2x) contains:
1. Taq PCR Master Mix (2x)
2. Water, nuclease free
3. 10 x Color Load
Taq PCR Master Mix (2x): Taq DNA Polymerase is supplied in 2 x Pol Buffer B containing 3 mM MgCl2 and 0.4 mM of each dNTP.
10 x Color Load: contains two gel tracking dyes and a gel loading reagent. It enables direct loading of PCR products onto an agarose gel.
Quality Control: All preparations are assayed for contaminating endonuclease, 3'-exonuclease, and nonspecific single- and double-stranded DNase activities. Typical preparations are greater than 95% pure, as judged by SDS polyacrylamide gel electrophoresis.
References:
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Chien, A., Edgar, D.B. and Trela, J.M. (1976) J. Bacteriol. 127, 1550.
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Kaledin, A.S., Sliusarenko, A.G. and Gorodetskii, S.I.(1980) Biokhimiya 45, 644.