title: GSM7876424: Mouse, Db 5 (ATAC-seq); Mus musculus; ATAC-seq
library_name: GSM7876424
library_construction_protocol: Briefly, a total of 20,000 cells per sample was washed twice with 50 µL of cold Dulbecco's Phosphate-Buffered Saline and re-suspended in 50 µL of lysis buffer. The suspension of nuclei was then centrifuged for 10 min at 500g at 4°C, followed by the addition of 50 µL of transposition reaction mix of TruePrep DNA Library Prep Kit V2 for Illumina. Samples were then incubated at 37°C for 30 min. DNA was isolated using the QIAquick PCR Purification Kit (QIAGEN, 28106). ATAC-seq libraries were first subjected to five cycles of pre-amplification using NEBNext High-Fidelity 2X PCR Master Mix (New England Biolabs, M0541S). To determine the suitable number of cycles required for the second round of PCR, the library was assessed by qPCR using NEBNext High-Fidelity 2X PCR Master Mix with SYBR Green I Nucleic Acid Gel Stain (Thermo Fisher Scientific, S7563) and then PCR amplified for the appropriate number of cycles. Libraries were purified with the QIAquick PCR Purification Kit.