title: GSM8282337: Arid1a_Cg1_ko_rep2; Mus musculus; OTHER
library_name: GSM8282337
library_construction_protocol: B cells were enriched by depleting 40LB cells after 4 days of co culture The epicypher CUT & Tag protocol was employed with slight modifications. Approximately 300,000 B cells obtained from 4 days of 40LB+ IL4 stimulation were used for CUT & Tag assay. Briefly, isolated nuclei from Cg1 Arid1a KO and control B cells were mixed with activated ConA beads and incubated for 10 minutes at room temperature. The nuclei-conjugated ConA bead complexes were incubated overnight at 4 ℃ with primary antibody Arid1a/BAF250A (1:50; cell signaling # 12354S) or control IgG (1:100; cell signaling #2729S) in digitonin 150 buffer. After primary incubation, nuclei-bead complexes were incubated with secondary antibody in digitonin 150 buffer for 1 hour at room temperature followed by the tagmentation step using CUTANA pAG-Tn5. Targeted chromatin tagmented products were column purified and libraries were amplified with 18 PCR cycles. Purified libraries were sequenced on Illumina NovaSeq 6000. Kaya-Okur, H.S., Wu, S.J., Codomo, C.A., Pledger, E.S., Bryson, T.D., Henikoff, J.G., Ahmad, K., and Henikoff, S. (2019). CUT&Tag for efficient epigenomic profiling of small samples and single cells. Nat. Commun. 10, 1930. https://doi.org/10.1038/s41467-019-09982-5.