MicroRNA, a class of non-coding RNAs with a length of about 22 nt, is widely concerned and plays a very important role in regulating gene expression in plants and animals. This kit uses the SYBR Green I chimeric fluorescence method for the quantitative detection of miRNA fluorescence. The 2×Hieff™ miRNA Universal qPCR Master Mix is a new generation of premix fluorescent quantitative PCR detection reagent specially developed for the quantitative detection of miRNA. It contains special ROX Passive Reference Dye. Suitable for most qPCR instruments, there is no need to adjust the concentration of ROX on different instruments, just add primers and templates in the preparation of the reaction system can be amplified. The DNA Polymerase uses chemically modified heat-activated Polymerase, combined with a special Buffer system, to make the reaction more specific, more sensitive, and can be accurately quantified in a wider range.This product is recommended for use with our Hifair™ miRNA 1st Strand cDNA Synthesis Kit (Tailing) (Cat# 11148) to obtain optimal experimental results. Features Platform-wide: No need to alter ROX concentration using premixed colors High sensitivity detection rate: 10 pg DNA can be detected Good amplification performance: Excellent amplification efficiency High specificity: Able to distinguish single base differences between miRNAs in the same family Applications Gene Expression Specifications Hot Start Built-in hot start Detection method SYBR PCR method qPCR Polymerase Taq DNA polymerase Type of sample DNA Application equipment Most qPCR instruments Product type SYBR premix for real-time fluorescence quantitative PCR Apply to (application) Gene Expression Components Components No. Name 11171ES03 11171ES08 11171-A 2×Hieff™ miRNA Universal qPCR Master Mix 1 mL 5×1 mL 11171-B RNase-free H2O 2×1 mL 5×1 mL Figures Figure 1. Good amplification performance: excellent amplification efficiency. miR-let-7b-5p, miR-let-7c-5p, miR-let-7e-5p, and miR-let-7f-5p genes were amplified using human 293T cells and mouse liver Total RNA as templates. The results showed that compared with similar products, Hifair™ miRNA 1st Strand cDNA Synthesis Kit (Cat# 11148ES) matched the reaction system of Hieff™ miRNA Universal qPCR SYBR Master Mix (Cat# 11171ES) with excellent performance. Figure 2. High specificity: be able to distinguish single base differences between miRNAs in the same family. The human hsa-let-7 family has multiple miRNAs, with few bases that differ from each other, or even only a single base difference. Amplify these miRNAs with different primers and use Formula 2- Δ CT x 100% calculated matching rate. The results showed that closely related subtype family members could be effectively distinguished.
