alias: GSM3473609
title: GSM3473609: Control - Poly(I:C) Exp #2 0 mins; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was purified from BMDMs using TRIzol reagent (Ambion) as per the manufacturer's instructions.  For fractionation experiments, confluent 10-cm dish of mature BMDMs were scraped into 400µL cold NP-40 lysis buffer, APJ1 (10mM Tris-HCl (pH 7.5), 0.08% NP-40, 150mM NaCl). Lysed cells layered onto 1mL cold sucrose 322 cushion, APJ2 (10mM Tris-HCl (pH 7.5), 150mM NaCl, 24% w/v sucrose) and centrifuged for 10min at 4°C and 13000 rpm. The supernatant from this spin represents the cytoplasmic RNA fraction, which is immediately added to 3 volumes of 100% ethanol and 2 volumes of buffer RLT (4M GuSCN, 325 0.1M β-mercaptoethanol, 0.5% N-lauroyl sarcosine, 25mM Na-citrate, pH7.2) and stored at -80°C until ready to purify RNA. Pellet, containing intact nuclei, is resuspended in 500µL TRIzol reagent. Full length cDNA libraries were barcoded using the Nextera XT Tagmentation protocol (Illumina).
identifiers.namespace: GEO
identifiers.id: GSM3473609