Features and Benefits - Saves preparation time by combining Precision™ Taq DNA Polymerase, dNTPs and
reaction buffer into a ready-to-use mixture.
- Reduces the risk of contamination by decreasing the number of pipetting steps.
- Provides consistent reaction performance and results.
The Precision™ Taq DNA polymerase is a thermostable product in abm’s PCR series
with outstanding fidelity due to its 3’ to 5’ proofreading activity. The newly introduced
2X PCR Precision™ MasterMix is a ready-to-use mixture containing abm’s Precision™
Taq DNA polymerase, dNTPs, and reaction buffer with proprietary additives in a 2X
concentration. It contains all the necessary reagents for reproducible, efficient and
high yield amplification of DNA. The 2X PCR Precision™ MasterMix with dye (Cat. No.
G124-dye) contains a proprietary blend of inert blue dye and stabilizer in precisely
balanced proportions. This allows easy, direct loading of the final PCR products onto a
gel for analytical electrophoresis.
abm’s Precision™ Taq DNA polymerase possesses excellent thermostability which
allows it to replicate DNA at 70°C - 80°C. It catalyzes the polymerization of nucleotides
into duplex DNA in the 5’ to 3’ direction in the presence of magnesium. The enzyme has
a molecular weight of approximately 90 kDa. Unlike Taq DNA Polymerase, Precision™
Taq DNA Polymerase exhibits 3’ to 5’ exonuclease (proofreading) activity, which
enables the polymerase to correct errors in nucleotide incorporation. As a results, the
error rate of Precision™ Taq DNA Polymerase is 2.6 x 10-6 per nucleotide per cycle.
The enzyme is ideal for PCR applications that demand high fidelity and can amplify
templates up to 6 kb in length. The amplified PCR products are blunt-ended.
Shipping and Storage Keep at -20°C for long term storage. 2X PCR Precision™ MasterMix and 2X PCR
Precision™ MasterMix with dye are stable at 4°C for one month or fifteen freeze-thaw
cycles. For daily use, it is recommended to keep an aliquot at 4°C.
Protocol 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. The use of
“clean”, automatic pipettors designated for PCR and aerosol resistant barrier tips are
recommended. Always keep the control DNA and other templates to be amplified
isolated from the other components.
A negative control reaction (omitting template DNA) should always be performed in
tandem with the sample PCR to ensure 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 |
| 2X PCR PrecisionTM MasterMix/ with dye | 25 µl | 1 X |
| Nuclease-free H2O | Up to 50 µl | - |
- We recommend preparing a mastermix for multiple reactions to minimize reagent loss and enable accurate pipetting
2. Mix contents of the tube and centrifuge briefly.
3. Incubate tube in a thermal cycler according to the following program:
| Step | Temp | Duration | Cycle(s) |
| Initial Denaturation | 94° C | 3 mins | 1 |
| Denaturation | 94° C | 10 secs | 30-40 |
| Annealing | 45 - 72° C | 30 secs |
| Extension | 72° C | 1 min/ 1 kb template |
| Final Extension | 72° C | 5 mins | 1 |
| Final Holding | 4° C | - | 1 |
*
The samples can be stored at -20°C until use. 4. Analyze the amplification products by agarose gel electrophoresis and visualize
by ethidium bromide or SafeView™ (Cat No. G108) staining. Use appropriate
molecular weight standards
