Analysis of Expression of Small Regulatory Rnas in Barley

Small RNAs (sRNA) of 15-30 nucleotides regulate various processes in eukaryotes. Regulatory sRNAs are produced through the RNA interference (RNAi) pathway from double stranded RNA (dsRNA). Dicer or dicer-like enzyme processes the dsRNAs into 15-30 bp dsRNAs, which in turn bind to the RNA-induced silencing complex (RISC) to regulate gene expression. We previously cloned sRNAs from barley. In this experiment, we determined the expression pattern of these sRNAs in different organs of barley (lemma, palea, awn, seed, leaf, leaf sheath, stem, and rachis) using northern blotting. Total RNA was extracted using a Trizol-based method. The quality of the extracted RNA was determined spectrophotometrically and by agarose-formaldehyde gel electrophoresis. Thirty μg of total RNA was separated on a denaturing polyacrylamide gel for northern blotting. Results of the northern analysis will be presented at the poster session.