Mir-X miRNA qRT-PCR TB Green Kits are complete systems for microRNA quantitation by qPCR from total RNA or purified small RNA samples. Each kit is complete and includes reagents for both:
- First-strand cDNA synthesis from miRNA and
- Quantitative PCR (qPCR) of miRNA
Simple, Single-Step cDNA Synthesis
Unlike other vendors' kits, these microRNA quantitation kits provide a simple, single-step protocol for the first-strand synthesis reaction. Mir-X uses an optimized mix of poly(A) polymerase and SMART MMLV Reverse Transcriptase to synthesize first-strand cDNA from your microRNA containing RNA sample.
Sensitive qPCR Using MicroRNA-Specific Primers
The cDNA is then amplified and quantified by qPCR using your microRNA-specific primer and our TB Green Advantage qPCR Premix. Multiple microRNA species, as well as the mRNA targets of the microRNAs, can be amplified from a single cDNA sample. The system is able to detect microRNAs down to 50 copies.
Highly Specific Detection of MicroRNA
To demonstrate the specificity of Mir-X miRNA quantitation we used a series of 8 highly similar synthetic Let7 miRNA variants. We first spiked each of the Let7 miRNAs into separate samples of yeast polyA+ RNA and generated cDNA using the Mir-X single-tube reaction. We then tested a panel of variant-specific primers with each cDNA sample to determine each primer’s ability to specifically and individually quantify the Let7 subtypes in the cDNA sample. Despite the high degrees of similarity among the variants and the primers, Mir-X qPCR was highly specific in detecting each Let7 variant.
Large Format qPCR Kits
The kits are available in economical, large-sized formats that provide 200 or 600 qPCR reactions, and each kit includes our Mir-X miRNA First-Strand Synthesis Kit and TB Green Advantage qPCR Premix.