title: GSM5553694: WT colonic macrophages [ATA-Seq]; Mus musculus; ATAC-seq
library_construction_protocol: Siglec-F(-)Ly6C(lo)CD64(+) colonic macrophages were sorted from male WT (two mice) and Tle4cKO (two mice) mice at the same time. Cells from mice of same genotype were pooled together. Nuclei were extracted as previously described (Buenrostro et al. 2013). DNA libraries were prepared as previously described (Buenrostro et al. 2013) by using a commercial kit (Vazyme, Nanjing, China). Nuclei were pelleted by centrifugation (4 ℃, 500 g, 30 min) and resuspended in 50 μL reaction mix (5 μL Tn5 transposase (Vazyme), 10 μL 5X TTBL buffer (Vazyme), and 35 μL nuclease-free water). The transposition reaction was incubated at 37 ℃ for 30 min. DNA fragments were purified using the Purification Kit (QIAGEN). Then, the libraries of DNA fragments were amplified by PCR (Vazyme) using the following PCR conditions: 72 ℃ for 3min; 98 ℃ for 30 s; thermocycling at 98 ℃ for 15 s, 60 ℃ for 30 s and 72 ℃ for 3 min (11 cycles); and 72 ℃ 5 min. Finally, the libraries were purified using 0.5X/1.2X DNA clean beads (Vazyme).