GSM4548320: C_4686_S31; Mus musculus; RNA-Seq
Brain tissue was homogenized in Trizol, and total RNA was isolated by chloroform extraction (Toni, L.S., et al., Optimization of phenol-chloroform RNA extraction. MethodsX, 2018. 5: p. 599-608.) The concentration and quality of total RNA samples was first assessed using Agilent 2100 Bioanalyzer. A RIN (RNA Integrity Number) of five or higher was required to pass the quality control. Then five hundred nanogram of RNA per sample were used prepare dual-indexed strand-specific cDNA library using TruSeq Stranded mRNA Library Prep Kit (Illumina). The resulting libraries were assessed for its quantity and size distribution using Qubit and Agilent 2100 Bioanalyzer. Two hundred pico molar pooled libraries were utilized per flowcell for clustering amplification on cBot using HiSeq 3000/4000 PE Cluster Kit and sequenced with 2×75bp paired-end configuration on HiSeq4000 (Illumina) using HiSeq 3000/4000 PE SBS Kit. A Phred quality score (Q score) was used to measure the quality of sequencing. More than 97% of the sequencing reads reached Q30 (99.9% base call accuracy).
GEO Accession
GSM4548320