GSM8181336: Amygdala, Rhesus Monkeys and Humans; Macaca mulatta; Homo sapiens; RNA-Seq
Single nuclei suspensions were made following minor modifications to 10x genomics demonstrated protocol: Nuclei Isolation from Cell Suspensions and Tissue for Single Cell RNA Sequencing. (CG000124 revE). Briefly, 1 ml of lysis buffer (10mM Tris-HCl pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% Nonidet P40 substitute, in nuclease free water) was added to each sample, homogenized 20x, and incubated on ice for 5 minutes. Samples were centrifuged at 850 RCF for 5 minutes at 4C. The resulting pellet was rinsed 3 times by resuspending in 1.5% BSA in PBS and centrifuged at 850 RCF for 5 minutes at 4C. Finally, the pellet was resuspended in 250ul 1.5% BSA in PBS and passed through 40µm Flowmi Cell strainer (Bel-art, H13680-0040). All barcoding and library preparation was performed by the UC Davis Genomics Core using the Single Cell Whole Transcriptome Kit (W01030, V1.3.1 Parse Biosciences) with strict adherence to manufacturer's instructions Libraries were prepared by the UC Davis Genomics Core and sequenced on a NovaSeq S4 300 150 bp pair-ended reads (PE150)