Description
Fast high-yield Taq master mix with loading dye
GenieTaq Rapid High Yield 2X Master Mix (Blue) is the flagship of the GenieTaq line: fast and robust. A dual-antibody hot-start and an optimised buffer give a high success rate and specificity at an extension rate of 15 sec/kb, and as fast as 1 sec/kb for fragments under 2 kb. Just add primers and template.
The polymerase is blocked by two antibodies that bind different epitopes, giving a high blocking rate. The mix amplifies genomic, plasmid and λDNA templates up to 15 kb and cDNA up to 10 kb, works across 24%–79% GC content and supports 7-plex amplification. A protectant keeps the mix from freezing at -20°C, so it is ready to use straight from storage, and the built-in tracking dye allows direct gel loading. Products carry 3'-dA overhangs for TA cloning and are compatible with the GenieClone Ultra DNA Assembly Kit and TOPO cloning kits. For research use only.
Key Features
| Rapid amplification | Extension at 15 sec/kb, and as fast as 1 sec/kb for fragments under 2 kb. | |
| Dual-antibody hot start | Two blocking antibodies give superior specificity and sensitivity. | |
| Broad compatibility | Works with 24%–79% GC templates and 7-plex amplification. | |
| Long targets | Amplifies genomic, plasmid and λDNA up to 15 kb and cDNA up to 10 kb. | |
| Ready to use | Stays liquid at -20°C and includes a tracking dye for direct gel loading. |
Specifications
| Format | 2X ready-to-use master mix with tracking dye |
| Hot-start technology | Dual-antibody (two epitopes) |
| Extension speed | 15 sec/kb (as fast as 1 sec/kb for fragments <2 kb) |
| Amplicon length | Up to 15 kb (genomic, plasmid, λDNA); up to 10 kb (cDNA) |
| GC range | 24%–79% |
| Templates | Genomic DNA, plasmid or virus DNA, cDNA and crude samples |
| Shelf life | 2 years |
Components
| Components | Amount |
| GenieTaq Rapid High Yield 2X Master Mix (Blue) | 5 × 1 ml / 15 × 1 ml / 50 × 1 ml |
Storage
Store at -30 ~ -15°C.
- Prepare the reaction system
- Set up a 50 μl reaction as follows:
| Components | Volume |
| ddH2O | Up to 50 μl |
| GenieTaq Rapid High Yield 2X Master Mix (Blue) | 25 μl |
| Forward primer (10 μM) | 1.5 μl |
| Reverse primer (10 μM) | 1.5 μl |
| Template* | x μl |
*The optimal amount depends on the template. Recommended amount in a 50 μl reaction:
| Template type | Amount |
| Genomic DNA | 10–200 ng |
| Plasmid or virus DNA | 50 pg - 50 ng |
| cDNA or crude samples | 1–5 μl (≤1/10 of the total PCR volume) |
- Run the reaction program
| Step | Temperature | Time | Cycles |
| Initial denaturation | 95°C | 2 min | 1 |
| Denaturation | 98°C | 10 sec | 28–35 |
| Annealing* | Tm | 30 sec | 28–35 |
| Extension | 68°C | 15 sec/kb | 28–35 |
| Final extension | 68°C | 5 min | 1 |
*Set the annealing temperature according to the primer Tm. If needed, optimise it with a temperature gradient. Specificity depends directly on the annealing temperature: raising it improves specificity.
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 the primer concentration; increase the extension time; increase cycles to 35–40; and use high-purity template.
Q2: How can I reduce nonspecific bands or smears?
- Optimise primer design; raise the annealing temperature in 2°C steps up to 65°C; decrease the final primer concentration to 0.2 μM; reduce the extension time if nonspecific bands are larger than the target; reduce cycles to 25–30; and use high-purity template.
Q3: Can I load the PCR product directly on a gel?
- Yes. The mix contains tracking dyes, so products can be analysed by electrophoresis directly after the reaction.
Q4: Which cloning methods are the products suited to?
- The products are compatible with the GenieClone Ultra DNA Assembly Kit and with TOPO cloning kits.
Q5: Does the mix freeze at -20°C?
- No. A protectant keeps the 2X master mix from freezing at -20°C, so it is ready to use straight from storage.
