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| Product Name | Hot-Start E-Taq DNA Polymerase (5 U/μL, High Sensitivity) |
| Catalog No. | IVDRM-0253 |
| Description | Hot-Start E-Taq DNA Polymerase (5 U/μL) is a dual-blocked hot-start Taq DNA polymerase. Developed through rational enzyme design and directed evolution—including structural analysis of DNA polymerases, multi-source sequence alignment, site-directed mutagenesis, random mutagenesis screening, stress testing and enzyme fragment recombination—an optimized Taq mutant was obtained and combined with a proprietary blocking antibody for dual blockage. The dual blockage inhibits both the 5'→3' polymerase activity and the 5'→3' exonuclease activity of Taq DNA polymerase. After 30 sec heating at the pre-denaturation temperature, the blocking antibody is completely inactivated, releasing both polymerase and exonuclease activities. This enzyme not only prevents non-specific amplification caused by mismatches or primer-dimers, but also suppresses fluorescence signal decay caused by probe degradation, doubly improving reagent specificity and ensuring greater stability during transport or room-temperature handling. Activation is unaffected by buffer pH or ionic strength, making it suitable for all hot-start PCR and qPCR reactions based on Taq DNA polymerase, particularly amplification of low-copy genes from complex templates (genomic DNA, cDNA). |
| Intended Use | Raw material enzyme for hot-start PCR and qPCR assays requiring high sensitivity and specificity; amplification of low-copy targets from complex templates such as genomic DNA and cDNA. |
| Principle / Technology | Dual antibody hot-start mechanism: the blocking antibody simultaneously blocks the 5'→3' polymerase and 5'→3' exonuclease activities of the engineered Taq DNA polymerase. A 30 sec incubation at the pre-denaturation temperature fully inactivates the antibody, releasing both activities. Activation is independent of buffer pH and ionic strength. |
| Detection Method | Hot-start PCR; qPCR; RT-qPCR |
| Sample Type | Complex templates including genomic DNA and cDNA; low-copy and low-concentration samples |
| Performance Range / Specifications | Strong specificity: dual-blocked hot-start enzyme doubly improves reagent specificity and stability. High sensitivity: limit of detection down to 5 copies/reaction. Excellent amplification efficiency: 99% efficiency with correlation R2 = 0.99 over a template range of 5×10^6–50 copies/reaction (demonstrated with an African swine fever target). Outstanding stability: a complete qPCR premix containing primers and probe remains stable after 14 days at 37 °C and after 50 freeze-thaw cycles; RT-qPCR systems formulated with this enzyme stably detect 4-plex respiratory targets after 14 days at 4 °C or 37 °C. No nuclease residue: free of exonuclease, endonuclease and RNase. |
| Sensitivity / LOD | Limit of detection down to 5 copies/reaction; stable detection and high fluorescence values for low-concentration samples. |
| Specificity | Dual antibody blocking prevents non-specific amplification from mismatches and primer-dimers and inhibits probe degradation, doubly enhancing specificity. |
| Reaction Conditions / Protocol | Fully activated after 30 sec at the pre-denaturation temperature; activation is independent of buffer pH and ionic strength. Compatible with standard hot-start PCR/qPCR cycling. |
| Storage Conditions | Store at -25 °C to -15 °C. |
| Shelf Life | 2 years when stored at -25 °C to -15 °C. |
| Package Specifications | 250 U; 500 U; 1000 U; 10000 U; 25000 U; 100000 U |
| Product Form | Liquid enzyme solution |
| Quality Control | Tested free of exonuclease, endonuclease and RNase contamination. |
| Concentration | 5 U/μL |
| Source / Origin | Engineered Taq DNA polymerase mutant obtained by rational design and directed evolution, recombinantly expressed. |
| Expiration Date / Stability | 2-year shelf life at -25 °C to -15 °C. Formulated primer/probe qPCR premixes remain stable for 14 days at 37 °C and withstand 50 freeze-thaw cycles without performance loss. |
| Compatibility | Suitable for all hot-start PCR and qPCR reaction systems based on Taq DNA polymerase, including multiplex (e.g., 4-plex) assays. |
| Application Notes / Precautions | Recommended for low-copy gene amplification from complex templates; activation is unaffected by buffer pH or ionic strength, ensuring robust performance across different buffer systems. |
For research use only, not for clinical use.
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