GSM7728803: E92_rep_1_scATAC-seq; Macaca mulatta; ATAC-seq
Transfer fresh frozen tissue sample to a pre-chilled 1.5 mL Eppendorf tube, add 500 μL of chilled Nuclei Lysis Buffer, and then transfer to a Douncer. Dounce the tissue with A and B pestle for complete homogenization, filter with a 40 μm strainer, and add 500 μL of chilled Nuclei Lysis Buffer. Mix gently and incubate on ice for 5 min. Centrifuge the nuclei at 500 x g for 5 min at 4°C. Remove supernatant without disturbing the pellet. Isolate pure nuclei using gradient centrifugation, and quantify the nuclei using Trypan blue staining on a hemocytometer. For single-nucleus multi-omics library construction, the isolated nuclei were resuspended with 500 μL diltuted nuclei buffer (1 x nuclei buffer (10X Genomics, PN-2000207), 1mM DTT, 1U/μL RNase inhibitor, nuclease-free Water). The nuclei were counted using Trypan blue staining on a hemocytometer, and 6,000 nuclei were loaded per lane. All libraries were constructed based on the user guide of Chromium Next GEM Single Cell Multiome ATAC + GEX kit (10X Genomics). Libraries were sequenced on the Illumina NovaSeq 6000 platform.