- Description
- Specifications
Catalog #: 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 72-78 °C and remains > 95% active following 1-hour incubation at 95 °C.
Synonyms:
Taq Plus DNA Polymerase, Taq Plus DNA, TaqPDNA.
Source:
Recombinant E.coli contains Thermus aquaticus polymerase gene.
Formulation:
20mM Tris-HCl (pH 8.0), 1mM DTT, 0.1mM EDTA, 100mM KCl, Stabilizers, and 50%glycerol.
Note:
All reagents, including Taq Plus, should be mixed immediately before use.
Storage:
Stable for 5 days at 10 °C, for longer period of time store at -20 °C.
Concentration:
5U/ µl.
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).
Optimization of DNA Synthesis:
It is important to ad the reaction components in the following order1. H2O.
2. 10 x reaction buffer.
3. dNTPs.
4. DNA template and primers.
5. Taq Plus.
Reaction Conditions:
DNA synthesis is performed in 100?l of mixture containing 20-200µm dNTPs, 0.3-1µm Promers, 0.1-0.250 mg of template DNA, 10 ?l 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 95? for 5 minutes and then cool to 40-68? for 5 minutes to allow the primers to anneal to the template DNA.
Unit Definition:
One unit of the enzyme catalyzes the incorporation of 10nmole of deoxyribonucleotides into a polynucleotide fraction in 30 min at 74 °C.
Usage:
Denovo Biotechnology's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
