GSM1688586: C4; Homo sapiens; ChIP-Seq
Neuronal nuclei from prefrontal cortex BA9 were NeuN+ labeled and fluorescence activated sorted (FACS) for subsequent H3K4me3 ChIP-seq. Briefly, approximately 300mg to 500mg of -80oC frozen grey matter dissected from BA9 was homogenized in 5ml lysis buffer and ultracentrifuged over sucrose solution at 24,400 rpm for two hours at 4oC. The resulting pellet was suspended in 550ul PBS, and immunolabeled with NeuN for 60 minutes while rotated in darkness. NeuN+ and NeuN- fractions were separated and collected by FACS. ChIP sequencing was performed for the NeuN+ fraction. Briefly, genomic regions with H3K4me3 modification were extracted by chromatin immunoprecipitation using anti-H3K4me3 antibody and further prepared into ChIP-seq library for sequencing. For each brain sample, 1 ug of DNA was used to construct sequencing libraries using Illumina’s TruSeq Sample Prep Kit, according to the manufacturer’s protocol (Illumina, San Diego, CA), and sequenced using 36nt single-end reads on Illumina’s GA II.
GEO Accession
GSM1688586