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. dUTP/UDG system: Preventing Airborne DNA Contamination. 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
