GSM6125857: siCtrl, Veh, IgG, Rep 3 [CV-IgG3]; Homo sapiens; OTHER
Following treatment, samples were processed for CUT&RUN using the CUTANA ChIC/CUT&RUN Kit (Epicypher, #14-1048; protocol from user manual version 2.0). Library prep was performed using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, #E7103) and NEBNext Multiplex Oligos for Illumina (NEB, #E6440), with changes made to the protocol to improve the capture of smaller TF fragments: the inactivation temperature after end-repair was changed to 50°C for 1 hour, the bead size-selection ratio in the adaptor cleanup step was increased to 1.75x, and the bead size-selection ratio in the PCR cleanup step was increased to 1.2x. Samples were sequenced by paired-end 150bp sequencing on an Illumina NovaSeq 6000 by the Next Generation Sequencing department at the UW-Madison Biotechnology Center.