GSM8392682: NIH12, sEOAD, snMultiome [GEX]; Homo sapiens; RNA-Seq
Samples were stored at -80 °C. The nuclei isolation protocol was modified based on a 10x Genomics protocol (CG000375). Briefly, 30 mg of frozen tissue was finely minced on dry ice to the smallest possible size, and then it was homogenized in homogenization buffer on ice within 20 minutes. The homogenized tissue was filtered through a 35 μm cell strainer. Small cell debris were removed by centrifuging at 300g for 3 minutes at 4 °C. The large cell debris was removed by gradient centrifugation using OptiPrep density gradient medium (Sigma, cat#D1556-250ML) at 7200g for 15 minutes at 4 °C. The nuclei pellet was resuspended in a 0.1x lysis buffer. After washing the isolated nuclei in the wash buffer, the nuclei were resuspended in 1x nuclei buffer before single-nuclei Multiome library construction. (UTHealth Cancer Genomic Core) single-nuclei Multiome sequencing; Single-nuclei libraries were constructed following the 10x Chromium Single Cell Multiome ATAC + Gene Expression protocol (CG000338).