MiRNA Isolation from Plants Rich in Polysaccharides and Polyphenols

Abstract

MicroRNAs (miRNAs) are a group of 18-22-nucleotide-long noncoding RNAs, which show wide array of roles in various biological and metabolic processes in both animals and plants. They are formed endogenously and have evolutionary conserved sequences. RNA quality is extremely important for sequencing miRNAs using next-generation sequencing platforms. However, isolation and quantification of miRNAs from plant samples are often technically difficult and recovery of miRNAs from total RNA might be problematic. Degradation of RNA weakens the true miRNAs present in the sample, hence it is crucial to use a protocol that effectively retains good integrity miRNAs. In this chapter, we outline few protocols that can be used to maximize the retrieval of good-quality miRNA and total RNA to be used in miRNA analysis for plants rich in polysaccharides and polyphenols.

Keywords: NGS; Next-generation sequencing; Plant; Polyphenol; Polysaccharide; RNA; Secondary metabolite; miRNA; miRNA isolation.