miScript SYBR® Green PCR Kit
For miRNA detection by real-time PCR using the miScript PCR System
The miScript SYBR Green PCR Kit is an integral component of the miScript PCR System for miRNA detection and quantification. The kit is based on the proven technology of QuantiTect SYBR Green Kits. cDNA generated by the miScript II RT Kit is used as the template for real-time PCR with the miScript SYBR Green PCR Kit and either PCR arrays (miRNome or Pathway-Focused miScript miRNA PCR Arrays) or appropriate assays (miScript Primer Assays for miRNA or other noncoding RNA, miScript Precursor Assays for precursor miRNA, or QuantiTect Primer Assays for mRNA).
The miScript SYBR Green PCR Kit, together with the components of the miScript PCR System, ensures sensitive, specific miRNA detection and quantification. The miScript PCR System provides exceptional sensitivity and a broad dynamic range. Amplification efficiencies are consistently high, enabling accurate expression analysis of multiple miRNAs using the ∆∆CT method (see figure miRNA expression in colorectal cancer). The high performance of the miScript PCR System ensures highly reproducible results between technical and biological replicates at the first attempt.
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.
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.
Reverse transcription is performed using the miScript II RT Kit. The resultant cDNA is used as a template for real-time PCR performed with the miScript SYBR Green PCR Kit, together with appropriate assays or arrays. This makes miRNA quantification and profiling a straightforward procedure, accessible to any lab with a real-time PCR cycler.
The miScript SYBR Green PCR Kit is used as part of the miScript PCR System for:
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