title: Dysregulation of miRNAs in patients with Huntington's Disease
design: Dysregulation of miRNAs in patients with Huntington's Disease
library_construction_protocol: A miRNA analysis on peripheral blood mononuclear cells (PBMCs) was performed by RNA-seq in 30 subjects, including 15 HD patients and 15 healthy controls. The subjects were recruited at the Oasi Research Institute - IRCCS (Troina, Italy).  PBMCs separation was performed using Ficoll-Paque (Ficoll Plaque PLUS–GE Healthcare Life Sciences, Piscataway, NJ, USA.Total RNA extraction from PBMC was performed using TRIzol reagent (TRIzol Reagent, Invitrogen Life Technologies, Carlsbad, CA, USA), according to the manufacturer's instructions. RNA was stored at −80 ◦C until further processing.RNA concentration and purity were evaluated using NanoDrop™ 2000/2000c (Thermo Fisher Scientific),whereas sample integrity was analyzed by TapeStation 4200 (Agilent Technologies) using RNA ScreenTape Assay. 2.7. Indexed libraries were prepared from 250 ng purified RNA using QIAseq miRNA Library Kit (Qiagen) according to the manufacturer's instructions. Libraries were quantified using the TapeStation 4200 (Agilent Technologies) and Qubit fluorometer (Invitrogen Co.), then pooled such that each index-tagged sample was present in equimolar amounts. 