alias: GSM2989216
title: GSM2989216: 24h + T3 rep3; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was isolated using RNeasy Plus Micro Kit (Qiagen) and was quantified and purity checked using a NanoDrop ND-1000 (Thermo Scientific, Waltham, MA, USA). RNA integrity was verified using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). Two hundred ng of total RNA were used with the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEW ENGLAND BIOLABS) (Ipswich, MA) to construct index-tagged cDNA libraries. Libraries were quantified using a Quant-iT™ dsDNA HS assay with the Q-bit fluorometer (Life Technologies, Carlsbad, California). Average library size and the size distribution were determined using a DNA 1000 assay in an Agilent 2100 Bioanalyzer. Libraries were normalized to 10nM using Tris-Cl 10mM, pH8.5 with 0.1% Tween 20. Libraries were applied to an Illumina flow cell for cluster generation (True Seq SR Cluster Kit V2 cBot) and sequence-by-synthesis single reads of 50 base length using the TruSeq SBS Kit v5 (Illumina) (Illumina, San Diego, CA) were generated on the HiSeq 2500 following the standard RNA sequencing protocol. For demultiplexing process, we used bcl2fastq with standard parameters and 1 mismatch allowed for the index identification. RNA-Seq was performed at the Genomics Unit of the Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid.
identifiers.namespace: GEO
identifiers.id: GSM2989216