title: GSM9654445: Ser2P-phosphorylated Pol II CUT&RUN on WTC11 iPSC-derived NPCs (20 days of differentiation); Homo sapiens; OTHER
library_name: GSM9654445
library_construction_protocol: NPCs were harvested using Accutase. The cell pellet was resuspended in 1 mL ice cold PBS (Gibco). The cells were centrifuged at 500g for 3 min at 4 °C and the cell pellet was resuspended in 1 ml of nuclei isolation buffer (NIB: 10 mM HEPES-KOH, pH 7.9, 10 mM KCl, 0.1% NP40, 0.5 mM spermidine and 1× Halt protease inhibitor cocktail) and incubated for 10 min on ice. Nuclear pellet was collected by centrifuging at 600g for 3 min at 4°C and again resuspended in 1 ml of NIB to wash. Nuclei were fixed in 0.1% formaldehyde for 2 minutes and washed in NIB. Fixed nuclei were bound to concanavalin A beads and ~100,000 nuclei were subjected to GO-CaRT/CUT&RUN. Sequencing libraries for GO-CaRT/CUT&RUN were generated using KAPA HyperPrep Kit (KAPA, KK8504) following the manufacturer's instructions. The libraries were amplified for 12–14 PCR cycles. DNA fragments were quantified by Tapestation D1000 (Agilent) and Qubit high sensitivity dsDNA (Invitrogen) assays, pooled, and sequenced by 150-bp paired-end sequencing on NovaSeq X Plus systems (Illumina).
identifiers.namespace: GEO
identifiers.id: GSM9654445_r1