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Hieff™ miRNA Universal qPCR Master Mix (SYBR) is a new-generation fluorescent quantitative PCR premix developed specifically for quantitative miRNA detection using the SYBR Green I intercalation method. MicroRNAs (~22 nt) are non-coding RNAs that play critical regulatory roles in gene expression. The formulation uses a chemically modified heat-activated Taq DNA Polymerase with specialized buffer, delivering high specificity, sensitivity, and accurate quantification across a wide dynamic range.
Contains universal ROX Passive Reference Dye compatible with most qPCR instruments without adjustment. Designed for use with Hifair™ miRNA 1st Strand cDNA Synthesis Kit (Cat# 11148ES).
Features
- Chemically modified Hot Start Taq: improved specificity and sensitivity
- Universal ROX Passive Reference Dye: compatible across most qPCR platforms
- Single-base discrimination: distinguishes closely related miRNA family members (e.g., hsa-let-7 family)
- Excellent amplification efficiency across diverse miRNA targets and species
- Wide quantification range
MicroRNAs are approximately 22 nucleotides in length — too short for standard Tm-based primer design used in mRNA qPCR. miRNA quantification requires specialized stem-loop RT primers (during cDNA synthesis) and matched universal qPCR conditions that can amplify the short stem-loop cDNA product with high efficiency and specificity. Standard qPCR master mixes are not optimized for these short amplicons.
ROX (6-carboxy-X-rhodamine) is a passive reference dye that normalizes non-PCR-related signal variation (instrument fluctuations, pipetting variation) by providing a stable fluorescence baseline. This mix contains a universal ROX concentration compatible with most qPCR instruments across brands without requiring ROX concentration adjustment.
Yes. Validated for single-base discrimination between closely related miRNA family members. For example, this mix can distinguish individual members of the hsa-let-7 family (let-7a, let-7b, let-7c, let-7e, let-7f), which differ by only 1–2 nucleotides.
This master mix is designed to be paired with the Hifair™ miRNA 1st Strand cDNA Synthesis Kit (Cat# 11148ES), which uses stem-loop RT primers for miRNA-specific first-strand cDNA synthesis.
Valid results require: amplification efficiency 90–110%; R² > 0.98 for standard curves; S-shaped amplification curve with Ct values between 15 and 35; and a single-peak melting curve. Ct values below 15 may cause threshold baseline issues. No-template control (NTC) Ct should be greater than 35.
Yeasen Biotechnology supports custom formulation and scale-up services for reagent mixes and master mixes. Contact BioHippo to discuss project requirements and initiate an inquiry.
▶ GMP-Grade & Bulk Supply
Select Yeasen reagent mixes are available in GMP grade for IVD assay development. Manufactured in an ISO 13485-certified UCF.ME™ facility with FDA Drug Master File (DMF) support.
- GMP release testing and CoA documentation
- Scalable from milliliter to liter-scale quantities
- Consistent lot-to-lot performance specifications
▶ Glycerol-Free & Lyophilization-Ready Formats
Glycerol-free and lyophilization-ready formats are available for reagents intended for cartridge-based point-of-care devices, liquid handling automation, or ambient-temperature distribution.
- Glycerol-free liquid formulation
- Lyophilization-compatible format with cryoprotectant
- Custom reaction buffer optimization
▶ Molecular IVD RDC Service
Yeasen's Research and Development Contracting team provides end-to-end support for molecular diagnostic product development using Yeasen reagent mixes as a foundation.
- Master mix selection and performance matching for target assay
- Primer/probe design and assay buffer optimization
- Sensitivity, specificity, and LOD validation
- Instrument compatibility assessment
▶ Custom Formulation
Custom formulation services are available for researchers requiring modifications to standard reagent mix compositions, including ROX level adjustment, dye substitution, stabilizer optimization, and buffer modification.
- ROX concentration customization (No / Low / High ROX)
- Reference dye and indicator dye options
- Buffer composition and pH optimization
- Scale-up to commercial quantities on confirmation
ⓘ Customization services fulfilled by Yeasen Biotechnology. Contact BioHippo for project scoping and pricing.
