GSM5602332: C20; Homo sapiens; RNA-Seq
Nuclei were isolated from frozen tissue, as per 10x Genomics' recommendations, using chilled, 0.1% Nonident P40 lysis buffer with gentle homogenization and washed. Gel-Bead in Emulsions (GEMs) were generated on the sample chip in the Chromium controller. Barcoded cDNA was extracted from the GEMs by Post-GEM RT-cleanup and amplified for 12 cycles. Amplified cDNA was fragmented and subjected to end-repair, poly A-tailing, adapter ligation, and 10X-specific sample indexing following the manufacturer's protocol. Libraries were quantified using Bioanalyzer (Agilent) and QuBit (Thermofisher) analysis. Libraries were sequenced using a 2x100PE configuration on a NovaSeq instrument (Illumina, San Diego, CA), targeting a depth of 50,000-100,000 reads per nucleus.