2X PCR TaqFast MasterMix 5.0 ml (200 Rxns) Product Description The TaqFast DNA polymerase is a uniquely engineered version of Taq DNA polymerase
which allows it to achieve rapid PCR amplification. This member of abm’s PCR
polymerase product series is the perfect solution to ultimate speed and dramatically
reduced reaction time. The newly introduced 2X PCR Precision™ MasterMix is a
ready-to-use mixture containing abm’s TaqFast 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
TaqFastMasterMix with dye (Cat. No. G280-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 TaqFast DNA polymerase is capable of an extension speed of 6 kb/min – 6
times faster than the regular Taq polymerase. This polymerase catalyzes the 5’ – 3’
synthesis of DNA. It possess 5’ – 3’ exonuclease activity and lacks 3’ – 5’ proofreading
exonuclease activity: making it the ideal choice for high through-put PCR. In addition,
template-independent “A” can be attached at the 3’ end of the PCR product which
can then be cloned into TA cloning vectors for downstream applications.
Features and Benefits - Saves preparation time by combining TaqFast 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.
Shipping and Storage
Keep at -20°C for long term storage. 2X PCR TaqFast MasterMix and 2X PCR TaqFast
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 TaqFast MasterMix/ with dye | 25 µl | 1X |
| 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 | 5 secs | 30-35 |
| Annealing | 45 - 72° C | 15 secs |
| Extension | 72° C | 10 secs/ 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.
