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ATAC-seq was conducted using the Hyperactive ATAC-Seq Library Prep Kit for Illumina (Vazyme, TD711), following the manufacturer's instructions with minor modifications to the previously described protocol50. Approximately 510^5 pachytene spermatocytes, isolated by the STA-PUT method, were washed with 500 L PBS, centrifuged, resuspended in 50 L cold lysis buffer, and incubated on ice for 10 minutes to isolate the nuclei. After centrifugation, the nuclei underwent transposition with a 50 L Tn5 transposase reaction mix at 37C for 30 minutes, which was stopped by adding stop buffer and incubating at room temperature for 5 minutes. Adapter-ligated DNA fragments were extracted and PCR-amplified with indexing primers (Vazyme, TD202). The amplified DNA was size-selected (200-800 bp) using VAHTS DNA Clean Beads (Vazyme, N411-01), and the DNA library's quantity were assessed using a Qubit4 Fluorometer and Bioanalyzer 2100 (Invitrogen). Libraries were sequenced on an Illumina platform by Genedenovo Biotechnology Co., Ltd (Guangzhou, China). |
ATAC-seq |
GENOMIC |
other |
PAIRED
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