scATAC-seq of mus musculus: the testicle of an adult male mouse
Nuclei were lysed, and the resulting nuclear suspension was adjusted to an optimal concentration for downstream experiments. The nuclear suspension was incubated with a transposase mixture, which penetrated the nuclei and selectively cleaved open chromatin regions, fragmenting genomic DNA. Simultaneously, sequencing primer adapters were ligated to the ends of the generated DNA fragments. Using a microfluidic system, individual nuclei were co-encapsulated with barcoded gel beads within oil droplets, forming Gel Bead-in-Emulsions (GEMs). Gel beads within GEMs were lysed to barcodes, followed by nuclear lysis to expose DNA fragments. Thermal cycling generated single-stranded DNA fragments tagged with P5 adapters and 10X barcodes, which underwent linear amplification. GEMs were disrupted to recover barcoded DNA fragments. PCR amplification P7 adapters to the fragment ends, completing the sequencing library preparation. Following quality control assessment, the validated libraries were sequenced on an Illumina platform.
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