Description
Taq PCR master mix with loading dye
GenieTaq 2X Master Mix (Blue) is the dye-containing format of the standard GenieTaq master mix. A blue tracking dye lets you load PCR products directly onto a gel, with no separate loading buffer. It is ready to use for routine PCR, colony PCR and genotyping.
The mix contains Taq DNA polymerase, dNTPs and an optimised buffer system. Just add primers and template: fewer pipetting steps improve throughput and reproducibility. Protective agents keep the 2X mix stable after repeated freeze–thaw cycles. Products carry a 3'-A overhang for direct TA 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 gives good amplification results quickly. | |
| Direct gel loading | The built-in blue tracking dye means PCR products go straight onto a gel. | |
| Simple setup | Just add primers and template to the 2X mix. | |
| Freeze–thaw stable | Protective agents keep activity stable after repeated freezing and thawing. | |
| TA-cloning ready | Products have a 3'-A overhang for cloning into T vectors. |
Specifications
| Format | 2X ready-to-use master mix with blue tracking dye (Taq DNA polymerase, dNTPs, optimised buffer) |
| Application | Conventional PCR |
| Templates | Animal and plant genomic DNA, E. coli genomic DNA, cDNA, plasmid DNA, λDNA |
| Extension speed | 60 sec/kb at 72°C |
| Product ends | 3'-A overhang |
| Shelf life | 2 years |
Components
| Components | Amount |
| GenieTaq 2X Master Mix (Blue) | 5 × 1 ml / 15 × 1 ml / 50 × 1 ml |
Storage
Store at -30 ~ -15°C.
- Prepare the reaction system
- Taq DNA polymerase has some activity at room temperature. Set up reactions on ice, then transfer to the thermal cycler to reduce non-specific amplification.
- Set up a 50 μl reaction as follows:
| Components | Volume |
| ddH2O | To 50 μl |
| GenieTaq 2X Master Mix (Blue) | 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 Adjust the annealing temperature to the primer Tm, generally 3 ~ 5°C below Tm. For complex templates, adjust the annealing temperature and extend the extension time.
Q1: Do I need to add loading buffer before running a gel?
- No. The mix contains a blue tracking dye, so PCR products can be loaded directly for electrophoresis after the reaction.
Q2: Can I clone the PCR products?
- Yes. Products have an adenine at the 3' end and can be cloned into T vectors. They are also compatible with seamless and TOPO cloning kits.
Q3: Why should I set up reactions on ice?
- Taq DNA polymerase has some activity at room temperature. Preparing reactions on ice reduces non-specific amplification during setup and gives more specific results.
