GSM2769730: Hoxa5 cKO pool 3; Mus musculus; RNA-Seq
Brainstems were collected after euthanasia of pups at P21. Tissues were  snap frozen in liquid nitrogen, and stored at −80 °C until RNA was extracted. RNA was isolated using TRI Reagent (Sigma Aldrich, T9424) and quantified using a NanoDrop ND-1000 fluorospectrometer (Thermo Scientific, Waltham, MA, USA). RNA from three brainstems was pooled, and three pools were used per genotype. Illumina truSeq stranded total RNA library prep kit with ribo-zero was used to prepare libraries from 1 µg of total RNA following Illumina’s protocol. Ribosomal RNA was depleted from total RNA. The remaining RNA was purified, fragmented, primed with random hexamers and reverse transcribed into first strand cDNA followed by second strand synthesis. A single A nucleotide was added to the 3’end of the fragments before ligation to multiple indexing adapters. DNA fragments with adapter molecules on both ends were enriched by PCR and purified with Ampure XP magnetic beads. Libraries were validated on Bioanalyser DNA 1000 chip and quantified by qPCR with the KAPA library quantification kit.
GEO Accession
GSM2769730