title: GSM8186577: pTh17, Clns1a, Crosslinked Cut and Run, Rep 1; Mus musculus; OTHER
library_name: GSM8186577
library_construction_protocol: Samples were immobilized to ConA beads and resuspended in ice cold antibody buffer. 1 ug primary antibody or IgG negative control was added into samples followed by incubation at 4°C overnight. On day 2, samples were washed with digitonin buffer, and digested with pAG-MNase. Reaction was stopped by stop buffer containing E.coli spike-in control for sample normalization. Samples were then incubated for 10 min at 37°C to release cleaved chromatin. Supernatant containing released chromatin was collected, and a final concentration of 0.1% SDS and 0.2 μg/μl proteinase K were added into each samples. Incubated samples overnight at 55°C. Purified DNA from supernatant using the CUTANA DNA purification kit and quantified DNA using Qubit fluorometer DNA library was prepared using CUTANATM CUT&RUN Library Prep Kit (SKU: 14-1001). The libraries were pooled equimolarly, and loaded on theIllumina NovaSeq X Sequencing Systemand run on 1 lane of a NovaSeq X+ 10B 300 Cycle Flowcell.