The miScript PCR System enables sensitive, specific miRNA quantification and profiling using SYBR Green-based real-time PCR. The miScript PCR System covers all the steps involved in conversion of RNA to cDNA and subsequent real-time PCR detection of miRNAs.
The miScript PCR System comprises:
- miScript II RT Kit — this kit enables simple, single-step cDNA synthesis. A single cDNA synthesis reaction can be used for detection of hundreds of miRNAs. The dual buffer system meets the distinctive needs of miRNA quantification using real-time PCR.
- miScript SYBR Green PCR Kit — this kit includes QuantiTect SYBR Green PCR Master Mix and the miScript Universal Primer, a reverse primer that allows detection of miRNAs in combination with a miScript Primer Assay or miScript miRNA PCR Array.
- miScript Primer Assay — miScript Primer Assays are miRNA-specific forward primers designed to detect mature miRNAs.
- miScript miRNA PCR Array — miRNome or Pathway-Focused miScript miRNA PCR Arrays are preformatted, single-use PCR arrays for rapid profiling of mature miRNAs.
- miScript miRNA PCR Array data analysis tool — this complimentary, Web-based tool simplifies the ΔΔCT method of relative quantification for miScript miRNA PCR Arrays.
Real-time PCR detection of miRNA
The miScript SYBR Green PCR Kit includes QuantiTect SYBR Green PCR Master Mix and the miScript Universal Primer, which is specific for the universal tag sequence added to the 5' end of the cDNA during reverse transcription. A single cDNA synthesis reaction is sufficient to interrogate hundreds of miRNAs. cDNA prepared using miScript HiSpec Buffer can be used for mature miRNA quantification with miScript miRNA PCR Arrays or miScript Primer Assays. cDNA prepared using miScript HiFlex Buffer can be used for detection of different RNA species with appropriate primers in the real-time PCR step: miScript Primer Assays for miRNA or other noncoding RNA, miScript Precursor Assays for precursor miRNA, or QuantiTect Primer Assays for mRNA.