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| Product Name | miRNA Universal qPCR SYBR Master Mix (Poly(A) Tailing Method) |
| Catalog No. | RESK-0011 |
| Description | MicroRNAs are a class of ~22 nt non-coding RNAs that play essential regulatory roles in gene expression in plants and animals. This kit quantifies miRNA by SYBR Green I intercalating dye-based fluorescent detection. The 2× master mix is a next-generation premix developed specifically for miRNA quantification. It contains a specially formulated universal ROX passive reference dye, making it compatible with all qPCR instruments without the need to adjust ROX concentration for different platforms—simply add primers and template to set up the reaction. A hot-start DNA polymerase combined with an optimized buffer system delivers superior specificity and sensitivity, enabling accurate quantification over a broad dynamic range. This product must be used together with a poly(A)-tailing miRNA first-strand cDNA synthesis kit to obtain final results. |
| Intended Use | For quantitative detection of miRNA by the poly(A) tailing (A-addition) method. Used in combination with a poly(A)-tailing miRNA first-strand cDNA synthesis kit for SYBR Green dye-based fluorescent quantitative PCR analysis of miRNA expression. |
| Principle / Technology | SYBR Green I intercalating dye chemistry for fluorescent quantitative PCR. A hot-start DNA polymerase paired with a proprietary buffer system enhances reaction specificity and sensitivity. A universal ROX passive reference dye is built into the mix, so no ROX concentration adjustment is required across different qPCR instruments. |
| Detection Method | Fluorescent quantitative PCR (SYBR Green dye-based) |
| Sample Type | cDNA synthesized from miRNA by the poly(A) tailing reverse transcription method; starting templates include total RNA or miRNA. |
| Performance Range / Specifications | Wide linear detection range. Sensitivity: 10 pg when total RNA is used as template; no fewer than 60 copies when miRNA is used as template. High specificity: distinguishes single-nucleotide differences among miRNAs of the same family. Instrument compatibility: universal for all platforms (No ROX, Low ROX, and High ROX systems). Stability: reagent stable for 7 days at 37°C; withstands 50 freeze-thaw cycles; assembled reaction mix remains stable for 48 h at 25°C without loss of performance. Amplification length: 60-300 bp; annealing temperature: 60°C; applicable GC range: 30%-70%. |
| Sensitivity / LOD | 10 pg total RNA; as few as 60 copies of miRNA. |
| Specificity | Effectively distinguishes single-base differences between closely related members of the same miRNA family (e.g., hsa-let-7 family subtypes). |
| Reaction Conditions / Protocol | Add primers and template cDNA to the 2× master mix; no ROX adjustment is required for any instrument. Annealing temperature: 60°C. Recommended amplification product length: 60-300 bp. For detailed cycling parameters, refer to the product manual. |
| Components / Formulation | 2× miRNA Universal qPCR SYBR Master Mix: 1 mL (100 rxn) or 5×1 mL (500 rxn) RNase-free Water: 2×1 mL or 5×1 mL |
| Storage Conditions | Store at -20°C protected from light. |
| Shelf Life | 1 year. |
| Package Specifications | 1 mL; 5×1 mL |
| Product Form | Mix |
| Concentration | 2× |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Valid for 1 year when stored at -20°C protected from light. The reagent tolerates storage at 37°C for 7 days and up to 50 freeze-thaw cycles; assembled reaction systems kept at 25°C for 48 h show no loss of performance. |
| Compatibility | Universal ROX formulation compatible with virtually all real-time qPCR instruments, including No ROX, Low ROX, and High ROX platforms—no ROX concentration adjustment needed. |
| Application Notes / Precautions | For the poly(A) tailing method, pair this product with a poly(A)-tailing miRNA first-strand cDNA synthesis kit for reverse transcription. Poly(A) tailing reverse-transcribes all miRNAs in the sample in a single reaction, so one reverse transcription product can be used to detect multiple miRNAs. Compared with the stem-loop method, the poly(A) method is simpler, but because all miRNAs are reverse-transcribed together, the risk of false-positive signals in subsequent qPCR is slightly higher. The stem-loop method offers sequence-specific reverse transcription and qPCR, but requires a stem-loop primer design and a separate reverse transcription reaction for each target miRNA. |
For research use only, not for clinical use.
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