AmpFAST 2X PCR SuperMix was premixed with an optimized concentration of AmpFAST Taq DNA Polymerase, dNTPs, Mg2+, and reaction buffer that could be used in extremely fast PCR experiments as well as massive gene detection requirements, which also performs better amplification functions for GC-rich and complicated secondary structures. For shorter sequence or plasmid samples, it is suggested to use 5 sec/kb elongation speed or less circuits to shorten your PCR reacting time. While for longer sequences (≥3 kb) or complicated samples that usually have lesser PCR products, it is suggested to use 15-30 sec/kb elongation speed or more circuits. Meanwhile, an “A” base at the 3’-end of the PCR products amplified by this enzyme can be directly used in TA cloning.
Kit Content(s)
AmpFAST 2X PCR SuperMix : 1 ml x 1 vial
Reaction Setup
| Component | Volume | Final Conc. |
| Forward primer, 10 μM | 2.5 μl | 0.5 μM |
| Reverse primer, 10 μM | 2.5 μl | 0.5 μM |
| AmpFAST 2X PCR SuperMix | 25 μl | |
| DNA template* | X μl | |
| PCR Grade Water | add to 50μ | - |
| Total volume: 50 μl | ||
| 2. Mix gently. If necessary, centrifuge briefly. Cap tubes and place in thermal cycler | ||
| 3. Process in thermal cycler for 25-35 cycles as follows: | ||
| Initial Denaturation | 2 mins at 95°C | |
| Denaturation | 15 secs at 95°C | 25-35 cycles |
| Annealing | 15 secs at 50-72°C | |
| Extension | 15 sec/kb at 72°C** | |
| Final extension: 5 min at 72°C | ||
Note: Optimal conditions for amplification will vary depending on the primers and thermal cycler used. It may be necessary to optimize the system for individual primers, templates, and thermal cyclers.