title: GSM8519342: TRIM66_WT_ChIP_seq_rep_2; Mus musculus; ChIP-Seq
library_name: GSM8519342
library_construction_protocol: Dissected MOE was dissociated by the Papain Dissociation System (Worthington) at 37 °C for 15 min and trituration for 5?15 times with a cut P1000 pipette tip. Cells were filtered by a 40- ?m strainer (Falcon). After spinning at 400×g for 2 min, cells were resuspended in DMEM (Gibco). ChIP-seq was performed on dis- sociated MOE cells with a ChIP-IT High Sensitivity Kit (Active Motif) and an S2 focused-ultrasonicator (Covaris) (settings: frequency sweeping, intensity = 3, duty cycle = 2%, cycles per burst = 200, 15 cycles of 1 min on and 1 min off). The final DNA product was eluted into ~60 uL. Libraries were prepared with an NEBNext Ultra DNA Library Prep Kit (New England Biolabs) and 11?18 cycles of PCR amplification, and then size-selected with PCRClean DX magnetic beads (Aline Biosciences) (0.55× to remove long fragments, and 0.9× to remove short fragments). Libraries were quantified by a Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) and a High Sensitivity DNA Analysis Kit on a Bioanalyzer 2100 (Agilent), and then sequenced on a MiSeq or a HiSeq 2500 (Illumina).
identifiers.namespace: GEO
identifiers.id: GSM8519342_r1