- Home
- IVD
- By Technology Types
- By Diseases Types
- By Product Types
- Research
- Resource
- Distributors
- Company
| Product Name | One-Step RT-qPCR Enzyme Master Mix |
| Catalog No. | IEC-HMM-0050 |
| Description | A ready-to-use enzyme mixture combining a thermostable reverse transcriptase with a hot-start DNA polymerase in an optimized buffer system. This formulation enables reverse transcription and quantitative PCR amplification in a single tube without intermediate handling, reducing contamination risk and hands-on time. |
| Intended Use | For one-step reverse-transcription quantitative real-time PCR (RT-qPCR) detection of RNA targets, including gene expression analysis, viral RNA quantification, and pathogen detection workflows. |
| Principle / Technology | Reverse transcriptase synthesizes first-strand cDNA from RNA template during the reverse transcription step (45-55 °C). Subsequently, the hot-start DNA polymerase is activated at high temperature and amplifies the cDNA target in the same tube using sequence-specific primers and fluorescent detection probes or intercalating dyes. |
| Detection Method | Real-time fluorescence detection using SYBR Green I dye or hydrolysis probes (TaqMan). Quantification by standard curve or comparative Ct method. |
| Sample Type | Total RNA, mRNA, or viral RNA extracted from cells, tissues, blood, serum, plasma, swab eluates, and other biological specimens. |
| Performance Range / Specifications | Linear dynamic range: 7-8 orders of magnitude (10 to 10⁷ copies). Amplification efficiency: 90-110%. R² >= 0.990 for standard curves. Detection of as few as 1-10 RNA copies. |
| Sensitivity / LOD | Detects <= 5 copies of target RNA per 25 uL reaction with 95% confidence under optimized conditions. |
| Specificity | Hot-start polymerase plus optimized buffer chemistry minimizes primer-dimer and non-specific amplification. Reverse transcriptase with reduced RNase H activity improves cDNA yield from structured RNA. |
| Reaction Conditions / Protocol | Reverse transcription: 50 °C, 10-15 min (single cycle). RT inactivation/initial denaturation: 95 °C, 2 min. PCR: 35-45 cycles of (95 °C 10 s, 60 °C 30 s). Total run time: ~60-90 min depending on cycle number. |
| Components / Formulation | 2x One-Step RT-qPCR Master Mix containing: hot-start DNA polymerase, thermostable reverse transcriptase, dNTPs (dATP, dCTP, dGTP, dTTP/dUTP), MgCl₂, reaction buffer with stabilizers and enhancers, ROX reference dye (optional variant). RNase-free water included. |
| Storage Conditions | Store at -20 °C protected from light. Once thawed, the 2x master mix may be stored at 4 °C for up to 2 weeks. |
| Shelf Life | 18 months at -20 °C from date of manufacture. |
| Package Specifications | 100 reactions (1 mL), 500 reactions (5 mL), 2,500 reactions (25 mL). Typically based on 25 uL final reaction volume. |
| Product Form | Viscous, clear liquid of pale yellow to light amber color. The color is from proprietary stabilizer components and does not affect performance. |
| Quality Control | Each lot tested for: RT efficiency (>= 90%), no-template control (NTC) signal, RNase and DNase contamination, linearity across 7-log concentration range, lot-to-lot consistency with reference RNA panels. |
| Key Features | Single-tube workflow minimizing contamination risk; broad dynamic range; compatible with SYBR Green and probe chemistries; includes optional ROX passive reference for instrument normalization; suitable for multiplex detection. |
| Purity | Enzymes purified to >= 95% homogeneity; master mix free of detectable RNase, DNase, and non-specific nuclease activities. |
| Concentration | 2x master mix concentrate. Each 25 uL reaction requires 12.5 uL of master mix. |
| Activity / Unit Definition | RT activity: >= 200 U/uL in reaction mix. DNA polymerase: hot-start activated, >= 2.5 U per 25 uL reaction. Combined activity optimized for balanced RT and PCR steps. |
| Molecular Weight | RT enzyme: ~75 kDa; DNA polymerase: ~94 kDa. These are formulation components; the master mix is a blend of multiple enzyme and buffer species. |
| Source / Origin | Both reverse transcriptase and DNA polymerase produced by recombinant E. coli fermentation. RT gene engineered for thermostability and reduced RNase H activity. Polymerase gene derived from thermophilic bacteria. |
| pH Range / Optimal pH | Reaction buffer pH 8.3-8.5 at 25 °C. Optimal for both reverse transcription and subsequent PCR amplification. |
| Shipping Conditions | Shipped on dry ice or with cold packs. Stable at ambient temperature for up to 72 hours during transit. |
| Expiration Date / Stability | 18 months at -20 °C. After first thaw, store at 4 °C and use within 2 weeks. Do not re-freeze thawed aliquots. |
| Regulatory / Compliance | ISO 9001 and ISO 13485 certified manufacturing. Suitable as IVD raw material with regulatory dossier support provided. |
| Compatibility | Compatible with most real-time PCR instruments including instruments from Bio-Rad, Applied Biosystems, Roche, Qiagen, and domestic Chinese qPCR platforms. ROX reference dye provided for instruments requiring passive normalization. |
| Recommended Buffer System | 2x master mix includes proprietary buffer with dNTPs, MgCl₂ (final 3-5 mM), stabilizers, and enhancers. ROX reference dye included in dedicated variant. UDG/dUTP carryover prevention chemistry available in Plus variant. |
| Application Notes / Precautions | Thaw master mix at 4 °C or on ice, protected from light. Prepare reactions on ice. Add template RNA last. For multiplex reactions, optimize primer and probe concentrations empirically. Use ROX variant for Applied Biosystems instruments (high ROX) or Agilent platforms (low ROX). |
| Batch-to-Batch Consistency | Lot-to-lot Ct variation <= 0.5 cycles across 7-log concentration range using standardized RNA control panel. Amplification efficiency variation <= +/- 5%. |
For research use only, not for clinical use.
|
There is no product in your cart. |
Copyright © 2026 Alta DiagnoTech. All rights reserved.
