GSM5494578: P1_cochlearHC_ATAC_rep2; Mus musculus; ATAC-seq
Atoh1 positive hair cells were FACS purified from the P1 cochlea. Cells were lysised in lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.5%NP-40) for 10 min on ice. After lysis, the nuclear lysis were incubate with Tn5 transposome and 5×TTBL (Vazyme Biotech, TD502) at 37 ℃ for 30 min. 5×TS was added to stop the tagmentation reaction. DNA was purified by phenol-chloroform extraction with in the presence of 40 ng carrier RNA followed by ethanol precipitation and then amplified by PCR Sequencing library was prepared using TruePrep DNA Library Prep Kit V2 for Illumina. PCR was preformed to amplified the library for 15-20 cycles according to the DNA amount. After the PCR reaction, libraries were size selected by 0.5×SPRI beads and purified with additional 1×SPRI beads.  The libraries were then sequenced on a NovaSeq sequencer.