title: GSM2905798: Donor1_GFPBATF_antiBATF; Homo sapiens; OTHER
library_construction_protocol: CUT&RUN was performed as described in (https://elifesciences.org/articles/21856), using anti-GFP (ab290), anti-BATF (sc-100974), and rabbit anti-mouse (ab46540) antibodies. Briefly, 6 million cells (30 million cells for anti-GFP CUT&RUN in GFP-BATF-containing cells) were collected and washed. Nuclei were isolated and incubated rotating with primary antibody (GFP or BATF) for 2 hours at 4C. BATF CUT&RUN samples were incubated an additional hour with rabbit anti-mouse antibody. Next, nuclei were incubated with proteinA-micrococcal nuclease (provided by the Henikoff lab) for one hour at 4C. Nuclei were equilibrated to 0C and MNase digestion was allowed to proceed for 30 minutes. Solubilized chromatin CUT&RUN fragments were isolated and purified. Paired end libraries were generated as described in (https://elifesciences.org/articles/21856). Briefly, purified CUT&RUN DNA underwent end repair followed by ligation to Illumina TruSeq HT dual-indexed adapters. Ligation reactions were followed by two successive Ampure XP bead-based size selections to remove adapter dimers. Libraries were amplified using the KAPA DNA polymerase library preparation kit protocol and amplifying for 14 cycles. Amplified libraries were cleaned up with two successive Ampure XP bead-based size selections to remove adapter dimers.
identifiers.namespace: GEO
identifiers.id: GSM2905798