Detaibio’s High-Fidelity PCR Master Mix is a 2× concentrated ready-to-use PCR solution, optimized for high-yield, high-accuracy amplification. In addition to dNTPs and Mg²⁺, the mix contains a next-generation, genetically engineered DNA polymerase with both 5'→3' polymerase and 3'→5' exonuclease (proofreading) activity. Proprietary enhancers further improve amplification performance. The mix generates blunt-end products suitable for downstream applications such as homologous recombination.
Key Features
- High fidelity – Accurate replication with proofreading activity
- High specificity – Optimized for clean amplification
- High yield – Robust performance across various templates
Reaction Setup
| Component | Volume (μL) |
|---|
| ddH2O | Up to 50 |
| 2× PCR Master Mix | 25 |
| Template | X |
| Forward Primer (10 μM) | 1 |
| Reverse Primer (10 μM) | 1 |
1.Thaw on ice before use and briefly centrifuge after mixing.
2.Recommended template amounts: DNA 50–150 ng, Plasmid 5–50 ng, cDNA 1–5 μL (≤10% of total volume).
Troubleshooting
| Issue | Possible Cause | Solution |
|---|
| No or weak product | Suboptimal primers | Redesign or optimize primers |
| Suboptimal annealing | Run temperature gradient optimization |
| Low primer/template concentration | Adjust concentrations accordingly |
| Inadequate extension | Extend elongation time |
| Insufficient number of cycles | Rerun the reaction with more cycles |
| Non-specific bands or smearing | Suboptimal primers | Reduce primer conc. or redesign |
| Suboptimal annealing | Increase and optimize temperature |
| Inadequate extension | Shorten extension time for larger non-specific bands; lengthen for smaller ones. |
| Excessive cycles | Reduce cycle number |
| Low template quality | Use higher-purity template |
Notes
- Use high-quality DNA templates.
- Avoid repeated freeze–thaw cycles to preserve enzyme activity.
- For research use only. Not for clinical diagnosis or therapeutic use.
