alias: GSM3681212
title: GSM3681212: 42d_A1-1_E1; Mus musculus; RNA-Seq
library_construction_protocol: Animals were transcardially perfused with ice cold Ringer's solution only. To preserve RNA quality, all following procedures were done rapidly to minimize time between animal sacrifice and tissue collection. Brains and spinal cords were dissected and the intermediate layer of spinal levels C5/C6 was dissected by crude trimming with blades und a dissection microscope. The spinal samples were snap frozen in liquid nitrogen and stored at -80 °C until further processing. Total RNA was prepared from all samples simultaneously by using the NucleoSpin® RNA XS Kit (Macherey-Nagel, Germany) under the protocols of the manufacturer with an additional TRIZol/chloroform extraction step due to the high lipid content of the spinal cord. The TruSeq RNA Sample Prep Kit v2 (Illumina) was used in the succeeding steps. Briefly, total RNA samples (100 ng) were ribo-depleted using Ribo Zero Gold (Epicentre) and then fragmented. The fragmented samples were reverse transcribed to cDNA, end-repaired and polyadenylated before ligation of TruSeq adapters containing the index for multiplexing. Fragments containing TruSeq adapters on both ends were selectively enriched by PCR.  Sequencing was performed on the Illumina HiSeq 4000 (single end, 125 bp) using the TruSeq SBS Kit v4-HS.
identifiers.namespace: GEO
identifiers.id: GSM3681212