title: GSM3082642: FASN Mutant 3 ON P14; Mus musculus; RNA-Seq
library_construction_protocol: Optic nerves were dissected and immediately frozen in liquid nitrogen. Samples were stored at -80° C until processed for RNA extraction. Total RNA was isolated with a Qiagen MiniKit according to manufacturer's protocols, quantity and quality determined with Nanodrop Lite (NanoDrop Technologies, Thermo Fisher Scientific), Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and Bioanalyzer 2100 (Agilent, Santa Clara, CA). The TruSeq Stranded mRNA Sample Prep Kit (Illumina, San Diego, CA, USA) was used in the succeeding steps. Briefly,poly-A containing RNA molecules were purified from the total RNA samples (100 ng) using oligo-dT attached magnetic beads. Isolated RNA was reverse-transcribed into double-stranded cDNA, with actinomycin added during first-strand synthesis. The cDNA samples were fragmented, end repaired and polyadenylated before ligation of TruSeq adapters. Fragments containing TruSeq adapters on both ends were selectively enriched with PCR. The quality and quantity of the enriched libraries were validated using Qubit 2.0 Fluorometer and Bioanalyzer 2100. The product is a smear with an average fragment size of approximately 360 bp. The libraries were normalized to 10 nM in Tris-Cl 10 mM, pH 8.5 with 0.1% Tween-20. Sequencing was performed on an Illumina HiSeq 4000 in single end 125 bp mode.
attributes.tag: GEO Accession
attributes.value: GSM3082642