Hieff Unicon™ Universal TaqMan Multiplex qPCR Mix (ROX) is a multiplex real-time qPCR premix utilizing the company's next-generation antibody-based hot-start Taq enzyme. The premix contains all universal components required for qPCR amplification, including dNTPs, Mg²⁺ ions, Taq DNA polymerase, and UDG enzyme, but does not include gene-specific primers or probes, which need to be designed and synthesized by the user. The reagent incorporates pre-mixed ROX reference dye, eliminating the need for separate addition and supporting both Low ROX and High ROX quantitative PCR instruments. For instruments that do not require ROX correction, the ROX-free qPCR Mix (Hieff Unicon™ Universal Superfast qPCR Mix (One Tube), Cat#16924) is recommended.

This product is suitable for various applications requiring ROX dye correction. Combined with gene-specific primers, probes, and templates, it enables qualitative and quantitative analysis, including residual DNA detection in intermediates, semi-finished products, and finished products in the biopharmaceutical industry. It can be used with the company's magnetic bead-based residual DNA sample preparation kit (Cat#18461ES/18462ES). Additionally, it serves as a core raw material for scientific research and in vitro diagnostics, suitable for developing differential detection kits for human bacterial, fungal, viral, and animal pathogen infections.

Feature

• High flexibility: Provides a high-quality qPCR reaction system with no fixed primer/probe constraints. Users can design and synthesize gene-specific primers and probes for different targets and projects, suitable for R&D, QC, and clinical applications.
• Cost-effective: Raw material-grade, high cost-performance solution that significantly reduces the overall cost of batch testing, method validation, and routine QC.
• Stable and universal system: Directed evolution hot-start polymerase with optimized buffer system delivers high sensitivity, excellent reproducibility, and non-specific amplification suppression. Compatible with mainstream qPCR instruments, with tested detection range, limit of quantification, and precision for various pharmaceutical applications including residual nucleic acid quantification, gene copy number detection, virus screening, and cell QC.
• Built-in ROX reference dye: Compatible with both Low/High ROX quantitative PCR instruments, effectively eliminating well-to-well fluorescence variation and delivering excellent amplification curve profiles.
• Ultra-fast amplification: Supports ultra-fast amplification protocols within 30 minutes, significantly reducing overall experimental time and providing a one-stop, high-performance qPCR core raw material solution for molecular diagnostics and research quantitative detection.

Figure

Case Study 1: Detection Range

Using the 16985 reagent with primers and probes from the 41308 and 41332 kits, standard curves were generated for E.coli DNA Control (30 fg/μL – 300 pg/μL) and CHO DNA Control (3 fg/μL – 30 pg/μL).

Figure 1. Standard curve (left) and amplification curves (right) of E.coli fragment using 16985.

Figure 2. Standard curve (left) and amplification curves (right) of CHO fragment using 16985.

Results: Amplification efficiencies were between 90–110% with R² > 0.99 for both E.coli and CHO DNA standards.

Case Study 2: Limit of Quantification

Using the 16985 reagent with primers and probes from the 41308 and 41332 kits, the limit of quantification was validated by testing E.coli (30 fg/μL) and CHO (0.3 fg/μL) standards in 10 replicates. 

Results: The limit of quantification for E.coli residual DNA and CHO residual DNA detection using the 16985 reagent reached 30 fg/μL and 0.3 fg/μL, respectively.

Storage

Store at -25°C to -15°C, valid for 1 year.

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