Description
Taq PCR master mix for routine PCR
GenieTaq 2X Master Mix is a ready-to-use formulation of standard Taq DNA polymerase with dNTPs and an optimised buffer. Just add primers and template. It is the everyday workhorse for routine PCR, colony PCR and genotyping.
The premixed format cuts pipetting, reduces contamination risk and improves throughput and reproducibility. Protective agents keep the mix active through repeated freeze-thaw cycles. Products carry a 3'-dA overhang for direct cloning into T vectors, and are also compatible with seamless and TOPO cloning kits. For research use only.
Key Features
| Classic, reliable performance | An optimised buffer system delivers good amplification results quickly. | |
| Convenient operation | The ready-to-use premix saves time and reduces contamination. | |
| Freeze-thaw stable | Protective agents keep the mix active after repeated freezing and thawing. | |
| TA cloning ready | Products carry a 3'-dA overhang for direct cloning into T vectors. |
Specifications
| Format | 2X ready-to-use master mix (Taq DNA polymerase, dNTPs, optimised buffer) |
| Application | Conventional PCR |
| Templates | Genomic DNA, cDNA, plasmid DNA and λDNA |
| Extension speed | 60 sec/kb |
| Product ends | 3'-dA overhang |
| Shelf life | 2 years |
Components
| Components | Amount |
| GenieTaq 2X Master Mix | 50 × 1 ml |
Storage
Store at -30 ~ -15°C.
- Prepare the reaction system
- Taq DNA polymerase is partly active at room temperature. Set up reactions on ice, then transfer to the thermal cycler to minimise nonspecific amplification.
- Set up a 50 μl reaction as follows:
| Components | Volume |
| ddH2O | To 50 μl |
| GenieTaq 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.
Q1: Why should I set up reactions on ice?
- Taq DNA polymerase has some activity at room temperature. Preparing the reaction on ice before moving it to the thermal cycler reduces nonspecific amplification during setup.
Q2: Can I clone the PCR products?
- Yes. Products carry a 3'-dA overhang and can be cloned directly into T vectors. They are also compatible with seamless and TOPO cloning kits.
Q3: How much template should I use?
- In a 50 μl reaction, use 0.1–1 μg animal or plant genomic DNA, 10–100 ng E. coli genomic DNA, 1–5 μl cDNA (≤1/10 of the reaction volume), 0.1–10 ng plasmid DNA or 0.5–10 ng λDNA.
