title: GSM8416054: CUT&Tag, peritoneal B1-a cell, male, FLAG; Mus musculus; OTHER
library_construction_protocol: CUT&Tag (Vazyme, TD904)  assay was conducted following the manufacturer's protocol. Briefly, B-1a cells were incubated with ConA Beads Pro at room temperature for 10 min. Rabbit anti-Bcl11a antibody (Abcam, #Ab191401, 1:200) was added and incubated with the cells overnight at 4°C, and further incubated with goat anti-Rabit IgG (H+L) secondary antibody (Vazyme, Ab208) at room temperature for 1 hour. After two cell washes, pA/G-Transposon Pro Enzyme was added and incubated with cells at 4°C for 1 h, followed by fragmentation with Mgcl2. CUT&Tag libraries of sorted B-1a cells were prepared using the Hyperactive Universal CUT&Tag Assay Kit for Illumina Pro (Vazyme, TD904), and qualified by using a Qubit 3.0 Fluorometer (Invitrogen, Carlsbad, CA, USA) and Agilent Bioanalyzer 2100 (Agilent Technologies, Palo Alto, CA, USA). The libraries with different indices were multiplexed and loaded onto an Illumina HiSeq 4000 instrument following the manufacturer's instructions (Illumina, San Diego, CA, USA). Sequencing was conducted using a 2 × 150 paired-end (PE) configuration. The antibodies used for CUT&Tag were rabbit anti-Bcl11a (Abcam, #Ab191401, 1:50), normal mouse IgG control (CST, 68860L, 1:50), and goat anti-Rabbit IgG (H+L) secondary antibody (Vazyme, Ab208)