title: GSM8368693: RepB_ES_d7_Sense,rep2; Mus musculus; OTHER
library_construction_protocol: For CHART-seq, 25 million cells underwent crosslinking with 1% formaldehyde, followed by additional crosslinking of nuclei with 3% formaldehyde. Chromatin was then sheared via sonication to produce fragments ranging from 2 to 4kb. Pre-clearing of the chromatin was performed using MyOne Streptavidin C1 beads. Subsequently, the pre-cleared chromatin was mixed with 36 pmol of either antisense (targeting Xist) or sense (used as a control) biotin-labeled probes, and captured using MyOne Streptavidin C1 beads. After washing the beads, DNA was eluted using RNase H. Eluted DNA was extracted via phenol-chloroform and further sheared to achieve fragments below 500 bp. The library was generated using the NEBNext® Ultra™ II DNA Library Prep Kit.