Description
High-yield Taq PCR master mix
GenieTaq High Yield 2X Master Mix delivers higher yield, stronger amplification and better fidelity than standard Taq DNA polymerase. It efficiently amplifies up to 10 kb genomic DNA fragments, 15 kb plasmid DNA fragments and 15 kb λDNA fragments, making it ideal for longer or higher-yield routine PCR.
The ready-to-use mix contains GenieTaq High Yield DNA Polymerase, dNTPs and an optimised buffer. Just add primers and template: fewer pipetting steps improve throughput and reproducibility. Protective agents keep the mix stable through repeated freeze-thaw cycles. It amplifies animal, plant and microbial DNA. Products carry a 3'-dA overhang for direct TA cloning and are also compatible with seamless and TOPO cloning kits. For research use only.
Key Features
| Higher fidelity and yield | Fidelity ~6× higher than Taq DNA polymerase, with higher amplification yield. | |
| Long targets | Amplifies up to 10 kb genomic DNA and 15 kb plasmid or λDNA fragments. | |
| Ready to use | Premixed formulation saves time and reduces contamination risk. | |
| Good stability | Performance remains stable after 50 freeze-thaw cycles. | |
| Cloning-ready products | 3'-dA overhangs allow direct TA cloning. |
Specifications
| Format | 2X ready-to-use master mix (DNA polymerase, dNTPs, optimised buffer) |
| Fidelity | ~6× higher than Taq DNA polymerase |
| Amplicon length | Up to 10 kb (genomic DNA), 15 kb (plasmid DNA), 15 kb (λDNA) |
| Templates | Animal, plant and microbial DNA |
| Product ends | 3'-dA overhang |
| Extension speed | 60 sec/kb |
| Shelf life | 2 years |
Components
| Components | Amount |
| GenieTaq High Yield 2X Master Mix | 50 × 1 ml |
Storage
Store at -30 ~ -15°C.
- Prepare the reaction system
- Set up a 50 μl reaction as follows:
| Components | Volume |
| ddH2O | To 50 μl |
| GenieTaq High Yield 2X Master Mix | 25 μl |
| Primer 1 (10 μM) | 2 μl |
| Primer 2 (10 μM) | 2 μl |
| Template DNA* | x μl |
*Recommended template amount in a 50 μl reaction:
| Template type | Amount |
| Animal and plant genomic DNA | 0.1–1 μg |
| E. coli genomic DNA | 10–100 ng |
| cDNA | 1–5 μl (≤1/10 of the total PCR volume) |
| Plasmid DNA | 0.1–10 ng |
| λDNA | 0.5–10 ng |
- Run the reaction program
| Step | Temperature | Time | Cycles |
| Initial denaturationa | 95°C | 3 min | 1 |
| Denaturation | 95°C | 15 sec | 30–35 |
| Annealingb | 60°C | 15 sec | 30–35 |
| Extension | 72°C | 60 sec/kb | 30–35 |
| Final extension | 72°C | 5 min | 1 |
a These conditions suit most reactions. For templates with complex structure, extend initial denaturation to 5–10 min.
b Set the annealing temperature 3–5°C below the primer Tm. For complex templates, adjust the annealing temperature and extend the extension time for efficient amplification.
Q1: What should I do if there is no amplification or the yield is low?
- Optimise primer design; run an annealing temperature gradient to find the optimum; increase primer concentration; extend the extension time; increase cycles to 35–40; use high-purity template; and decrease the amount of crude samples, or increase other templates within the recommended range.
Q2: How can I reduce nonspecific products or smeared bands?
- Optimise primer design; raise the annealing temperature towards 65°C in 2°C steps; decrease the final primer concentration to 0.2 μM; shorten the extension time if nonspecific bands are larger than the target; reduce cycles to 25–30; use high-purity template; and adjust the template amount to the recommended range.
Q3: Can I clone the PCR products directly?
- Yes. Products carry a 3'-dA overhang and can be cloned into T vectors. They are also compatible with seamless and TOPO cloning kits.
Q4: How should I adjust conditions for complex templates?
- Extend initial denaturation to 5–10 min, adjust the annealing temperature and extend the extension time.
