title: GSM4997989: ES AKO; Mus musculus; ATAC-seq
library_construction_protocol: 50,000 cells were harvested and lysed in 50 μl of cold lysis buffer (10 mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2, 0.1% (v/v) Igepal CA-630) for 10 min on ice to prepare the nuclei. Then, spin down immediately at 500 g for 5 min, 4 °C, to remove the supernatant. Nuclei were then incubated with TruePrep Tagment Enzyme (TD501, Vazyme) at 55 °C for 10 min. Immediately after the tag mentation, 1× VAHTS DNA Clean Beads (N411, Vazyme) was added into the reaction to purify DNA fragments. Nuclei were then incubated with TruePrep Tagment Enzyme (TD501, Vazyme) at 55 °C for 10 min. Immediately after the tag mentation, 1× VAHTS DNA Clean Beads (N411, Vazyme) was added into the reaction to purify DNA fragments. PCR amplify the library with the following program: 72 °C for 3 min; 98 °C for 30 s; and thermocycling at 1 cycle of 98 °C for 15 s, 15 cycles of 60 °C for 30 s and 72 °C for 3 min; following by 72 °C for 5 min. Libraries were purified with the 0.6×/0.15× double sided size selection. Libraries were sequenced with the Illumina HiSeq 2500 System.