GSM8783867: Input_WT_CC30_H3K27ac_rep2_H3K27me3_rep2_H3K4me3_rep2_HC; Mus musculus; ChIP-Seq
To perform ChIP-seq, we extracted  nuclei from hippocampus and brainstem of viral déjà vu mice at different time points after intravenous wtLCMV challenge, chromatin was fixed then we FAN-sorted neuronal nulclei.  Each biological replicate represents 200K  neuronal nuclei.fixed chromatin was incubated with proper ChIP-seq grade antibody, then DNA was extracted. non ChIPed genomic DNA was extracted to serve as input Nuclei were pelleted and resuspended with 25 µl of complete lysis buffer (lysis buffer + 1% protease inhibitor cocktail 200X provided in kit) per 100K nuclei and incubated for 5 minutes to ensure lysis then 75 complete Hank's balanced salt solution buffer (HBSS) (ThermoFisher Scientific, 14175095) (HBSS + 1% protease inhibitor cocktail 200X) was added per 100 K nuclei. Chromatin was sheared by sonication using a Picoruptor® (Diagenode) for 6 cycles. 20% of sample was set aside to serve as input and volume corresponding to 200K was subjected to magnetic immunoprecipitation using ChIP-seq grade antibodies: anti-H3K4me3 antibody (Diagenode, C15310003), anti-H3K27me3 antibody (Diagenode, C15410195-10), and anti-H3K27ac antibody (abcam, 4729). The ChIPmentation libraries were sequenced using 50 bp single-end reads on the HiSeq X (Illumina).