miScript Plant miRNA Solutions
For SYBR® Green-based real-time PCR profiling of plant miRNAs using the miScript PCR System
Plant miRNAs play a necessary role in processes such as development, disease resistance and stress response. These small non-coding RNAs are thought to regulate gene expression in plants at the post-transcriptional level by targeting mRNAs for cleavage. These targeted alterations in gene expression are mediated by differential expression of hundreds of mature miRNAs that each bind to specific sequences in the target mRNAs. Therefore, profiling to coordinate regulation and expression of mature plant miRNAs provides invaluable insight into mechanisms that an organism is using to develop tissue, protect itself from drought or fend off disease. Until now, universal profiling of mature plant miRNAs by qRT-PCR was virtually nonexistent because polyadenylation-based reverse transcription reactions occur with very low efficiency due to the biochemical nature of plant miRNAs. QIAGEN has released a unique solution that allows universal reverse transcription of all mature plant miRNAs in a tissue. Learn how profiling miRNAs in plants can accelerate your research today!
miScript plant miRNA quantification: go where other RT reactions cannot
Typical RT kits cannot be used to quantify plant miRNAs. Small RNAs possessing a 2’-O-methyl (2’-O-Me) modification on their 3’ terminal base, such as plant mature miRNAs and piwi-interacting RNAs (piRNAs), are refractory to polyadenylation. As a result, they cannot be efficiently reverse transcribed using a polyadenylation-based cDNA synthesis approach, which is the underlying mechanism of various miRNA reverse transcription kits including the miScript II RT Kit. The miScript Plant RT Kit has been designed and optimized for real-time PCR-based small RNA expression analysis together with miScript SYBR Green PCR Kit and miScript miRNA PCR Arrays or miScript Primer Assays. These functionally verified assays and arrays are available for rice, corn, soybean, tobacco and other significant species in agriculture and plant research. The specialized miScript Plant RT Kit overcomes this by first ligating an adaptor to the 3′ terminus of plant miRNAs. Unlike polyadenylation, this ligation reaction is not negatively inhibited by the presence of a 2′-O-Me modification on the 3′ terminal base of plant miRNAs. Reverse transcription can be performed following ligation.
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