Taq Plus DNA Polymerase (PCR) 200 ul (1000 U) Product Description Taq Plus DNA Polymerase is a mixture of Taq DNA Polymerase and proofreading DNA
Polymerase, which allows for the amplification of DNA templates with high fidelity. The
two enzymes act synergistically during PCR to generate more accurate and longer
PCR products with greater yields compared to Taq DNA Polymerase alone. PCR
products contain a mixture of blunt ends and single base (A) 3’ overhang. The error
rate of this PCR amplification is 7.5 x10-6 per nucleotide per cycle.
The products can be used for direct T/A cloning, but its efficiency is not as high as PCR
products amplified with Taq polymerase alone.
| Product Components | 250 U | 1000 U |
| Taq Plus DNA Polymerase (5 U/µl) | 50 µl | 200 µl |
| 10X PCR buffer, with Mg2+ | 1 ml | 3 ml |
| 25 mM MgSO4 | 1 ml | 1 ml |
Storage Buffer Components 50 mM Tris-HCl (pH 8.0), 100 mM NaCl, 0.1 mM EDTA, 5 mM DTT, 50 % glycerol and 1.0 %
Triton®X-100.
Unit Definition One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides
into a polynucleotide fraction in 30 mins at 74°C.
Shipping and Storage Upon arrival, Taq Plus DNA Polymerase should be stored at -20°C. Avoid repeated
freeze-thaw cycles of all Taq Plus components to retain maximum performance. All
Taq Plus components are stable for 1 year from the date of shipping if stored and
handled properly.
Protocol The following basic protocol serves as a general guideline and starting point for any
PCR amplification. Optimal reaction conditions (incubation times, temperatures,
concentration of Taq DNA Polymerase, primers, MgSO4 and template DNA) may vary
and need to be optimized for each specific PCR.
All PCR experiments should be assembled in a nuclease-free environment. In addition,
DNA sample preparation, reaction set-up and subsequent reaction(s) should be
performed in separate areas to avoid cross contamination.
A negative control reaction (omitting template DNA) should always be performed in
tandem with sample PCR to confirm the absence of DNA contamination.
1. Add the following components to a sterile 0.2 ml PCR tube sitting on ice.
| Components | Volume | Final Concentration |
| Template DNA | 100 ng | 2 ng/µl |
| Forward primer (10 µM) | 1 - 2.5 µl | 200 - 500 nM |
| Reverse primer (10 µM) | 1 - 2.5 µl | 200 - 500 nM |
| 10X PCR buffer, with Mg2+ | 5 µl | 1X |
| 25 mM MgSO4 (optional)* | 0 - 3 µl | 1.5 - 3 mM |
| dNTP Mix (10 mM) | 1 µl | 200 µM |
| Taq Plus DNA Polymerase (5 U/µl) | 0.5 - 1 µl | 2.5 - 5 U |
| Nuclease-free H2O | up to 50 µl | - |
* Optimal Mg
2+ concentration is specific to each DNA template-primer set and
can only be determined experimentally.
• We recommend preparing a mastermix for multiple reactions to minimize
reagent loss and enable accurate pipetting.
2. Mix contents of tube and centrifuge briefly.
3. Incubate tube in a thermal cycler at 94°C for 3 mins to completely denature the
template.
4. Perform 30 - 35 cycles of PCR amplification as follows:
Denature: 94°C for 30 sec
Anneal: 45 - 72°C for 30 sec
Extend: 72°C for 1 min/1 kb template
5. Incubate for an additional 5 mins at 72°C and maintain the reaction at 4°C. The
samples can be stored at -20°C until use.
6. Analyze the amplification products by agarose gel electrophoresis and visualize
by ethidium bromide or SafeView™ (Cat No. G108) staining. Use appropriate
molecular weight standards.
