Pro Multiplex Probe qPCR Mix

Pro Multiplex Probe qPCR Mix is a 2X Master Mix designed for multiplex real-time quantitative PCR using probe-based detection. The reaction can be performed simply by adding template, primers, and probes. This product contains a mutant hot-start enzyme, Hotstart Taq DNA polymerase, modified with antibody technology, combined with Dongsheng's specially formulated PCR Buffer, enabling highly sensitive multiplex qPCR. The reagent incorporates a dUTP/UDG contamination prevention system, which can eliminate dUTP-containing PCR products before amplification, effectively avoiding quantitative interference caused by cross-contamination of amplification products. This product is convenient for developing fully pre-mixed Master Mix molecular diagnostic kits with probes and primers. It is compatible with various fluorescent probes such as TaqMan and works seamlessly with common quantitative PCR instruments from manufacturers like ABI, Roche, and Bio-Rad. This product allows reaction setup without an ice box—the PCR Mix, primers, and template can be prepared at room temperature, and the prepared PCR reaction system can be stored at room temperature for 24 hours without loss of amplification efficiency.

 Applications

• Probe gene expression analysis  

• Probe Low-copy gene detection

• Multiplex probe qPCR 

• IVD diagnostic kit research

 Features

• Ready-to-use 2X Mix for multiplex qPCR

• High sensitivity and multiplex capability

• Built-in contamination control (dUTP/UDG)

• Broad probe and instrument compatibility

• Stable at room temperature for 24 hours

Fast Amplification Mode Supported: Run Time <50 min, Controlled ∆Ct Loss: ~1 cycle

Fast Amplification Testing:
Side-by-side testing evaluated P2731 and P2701 (probe-based qPCR mixes) under Standard (S, ~70 min) and Fast (F, ~48 min) cycling conditions. While both mixes showed an identical, minimal ∆Ct(F-S) of ~1.0 in the CY5 and FAM channels, they diverged significantly in the ROX channel. Driven by high thermal ramp rates that challenge probe hybridization and fluorescence capture, the ROX channel ∆Ct(F-S) jumped to 3.6 for P2701, but remained tightly regulated at 2.3 for P2731. These findings prove that P2731 delivers exceptional kinetic buffering to effectively mitigate fast-cycling efficiency losses, offering superior system tolerance and outperforming standard mixes in fast-program compatibility.

Target Allocation Strategy (Recommendation): In multiplex PCR assays, optimize channel assignment to offset fast-cycling kinetic losses:

ROX Channel: Allocate high-abundance, highly expressed, or non-LoD (Limit of Detection) target genes.

FAM or CY5 Channels: Reserve for fragile, ultra-low copy targets. These channels remain virtually unaffected by fast protocols ( ∆Ct ~1).