Certificate of Analysis and Data Sheet

Source:
Recombinant E.coli contains Thermus aquaticus polymerase gene.

Catalog No.
ENZ-309

Description:

Taq Plus is a mixture of Taq and Pfu. Taq Plus, which is used to improve the reliability and yield of conventional primer extension reaction. Taq Plus has two following advantages over Taq: (1) high fidelity with an error frequency 1.6/106 (or 0.0016/103) during DNA synthesis. (2) Taq Plus increases the efficiency of polymerization reaction, resulting in a great percentage of extenuation reaction completion up to 10 kb to 30 kb. Pfu has a temperature optimum between 72-78⁰C and remains > 95% active following 1-hour incubation at 95⁰C.

Unit Definition:

One unit of the enzyme catalyzes the incorporation of 10nmole of deoxyribonucleotides into a polynucleotide fraction in 30min at 74⁰C.

Storage Buffer:

20mM Tris-HCl (pH 8.0), 1mM DTT, 0.1mM EDTA, 100mM KCl, Stabilizers, and 50% glycerol.

Concentration:

2.5u/痞

10X Reaction Buffer:

200 mM TrisHCI (pH 8.8)
100 mM KCI
100 mM (NH4)2SO4
20 mM Mg SO4
1% Triton X-100
1 mg/ml bovine serum albumin (BSA).
Note: All reagents, including Taq Plus, should be mixed immediately before use.

Reaction Conditions:

DNA synthesis is performed in 100μl of mixture containing 20-200μM dNTPs, 0.3-1μM Promers, 0.1-0.250mg of template DNA, 10 μl of 10 x reaction buffer and 2.5-5 units of Taq Plus. Mix the reaction gently, centrifuge briefly and then overlay with light mineral oil.
Initially, denature the reaction by incubating at 95⁰C for 5 minutes and then cool to 40-68 ⁰C for 5 minutes to allow the primers to anneal to the template DNA.

Optimization of DNA Synthesis:

It is important to ad the reaction componennts in the following order :
1. H₂O
2. 10 x reaction buffer
3. dNTPs
4. DNA template and primers
5. Taq Plus

Storage Conditions :

Stable for 5 days at 10⁰C, for longer period of time store at -20⁰C.
Please avoid freeze-thaw cycles.

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

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