Beskrivning
Hifair™ Universal Advanced Multiplex One-Step RT-qPCR Mix (UDG Plus) is a ready-to-use master mix designed for multiplex quantitative PCR directly from RNA templates. The reverse transcription and qPCR amplification occur in a single tube, streamlining workflow and minimizing the risk of cross-contamination. This reagent is optimized for multiplex RNA amplification
Features
- Broad compatibility: Robust performance with various primer–probe sets.
- Fast cycling: Enables rapid protocols with total run times ≤35 min.
- High specificity and sensitivity: Proprietary buffer minimizes non-specific amplification and enhances multiplex efficiency.
- Anti-contamination system: dUTP/UDG system prevents carryover contamination from previous PCR products.
- Sample versatility: Tolerant to complex samples, allowing direct RT-qPCR from swab specimens without RNA purification.
Components
|
Components No. |
Name |
16930ES60(100 T) |
16930ES80(1,000 T) |
16930ES92(10,000 T) |
|
16930-A |
5×Hifair™ MP Buffer |
500 μL |
5 mL |
50 mL |
|
16930-B |
Hifair™ Enzyme Mix |
100 μL |
1 mL |
10 mL |
|
16930-C |
GC-Enhancer |
500μL |
5 mL |
50 mL |
[Note]: 5× Hifair™ MP Buffer is an abbreviation for 5× Hifair™ Universal Advanced Multiplex One-Step RT-qPCR Buffer.
Hifair™ Enzyme Mix contains thermostable reverse transcriptase, HotStart Taq DNA polymerase, RNase inhibitor, and UDGase.
Storage
This product should be stored at -25℃~-15℃ for one year.
Specifications
|
Format |
Tube, Liquid |
|
Sample Type |
Total RNA, Viral DNA, Viral RNA, mRNA |
|
Detection Method |
Primer-probe |
|
Inhibitor Tolerance (Simple) |
High, supports direct RT-qPCR using oral swab samples with lysis buffer. |
|
Instrument Compatibility |
Compatible with mainstream qPCR instrument platforms. |
|
Precision |
CV of Ct values at high, medium, and low template concentrations ≤ 1%. |
|
Linearity Range |
5 copies/reaction – 5 × 10⁵ copies/reaction; R² > 0.99; 90% ≤ Efficiency ≤ 110%. |
|
Sensitivity |
Detects ≤ 50 fg total human RNA. |
|
Stability |
1. Freeze–thaw stability: ≥10 cycles (−20 °C freeze, normal thaw). 2. Accelerated stability: stable for 7 days at 37 °C and 14 days at 4 °C. 3. Semi–premixed system (with primers/probes): stable for 7 days at 37 °C. |
Application
Microbial Detection, RT-qPCR Detection; Diagnostic pathogen detection (respiratory tract, intestinal tract, etc.); animal pathogen detection (non-swine fever, etc.); food testing (microorganisms, etc.)
Figure
1. High sensitivity
Figure 1. Sensitivity test.
Low-concentration plasmid or T7 RNA templates were amplified (20 replicates). 16930 showed higher detection rate and stronger fluorescence than Hifair™ V2 Multiplex One Step RT-qPCR Probe Kit (Yeasen 16630,2G). For 50 fg human RNA, all 20 wells were detected. Sensitivity outperformed 16630 and supports rapid cycling.
2. Precision
Figure 2. Precision assessment.
Amplification was performed using high (10⁴ copies/μL), medium (10² copies/μL), and low (10 copies/μL) template concentrations in 20 replicates. Criteria: CV of Ct ≤0.8% (high/medium) and ≤1.5% (low). Targets: human rhinovirus (HRV-A), respiratory syncytial virus (RSV-A), and parainfluenza virus type II/III. Results: CV values for all concentrations were <5%, demonstrating excellent assay precision.
3. High Inhibitor Tolerance
Figure 3. Inhibitor tolerance testing
Oral swab lysates were used as templates. Compared with TE controls(TE buffer–diluted templates served as inhibition-free controls), Ct differences were within ±1 and Rn within 10%. 16930 maintained stable performance in multiplex detection from direct swab samples.
4. Superior amplification of high-GC targets
Figure 4. Amplification performance with high-GC targets.
For targets with 70% GC content, the 16930 reagent has superior performance with Hifair™ V2 Multiplex One Step RT-qPCR Probe Kit (Yeasen 16630,2G). The addition of GC Enhancer further improved amplification efficiency and signal strength.
5. Supports fast protocols
Figure 5. Compatibility with rapid cycling program.
Under the SLAN 48S 25-min fast cycling protocol, Ct values differed by less than ±0.5 compared with the conventional program, confirming support for rapid amplification. Targets included high-concentration human 293 RNA and low-concentration pathogen RNA templates.
Documents:
Safety Data Sheet
Manuals
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