GSM4120427: Human Entorhinal Cortex of aged Healthy individuals Ct1-Ct2; Homo sapiens; RNA-Seq
Entorhinal cortex tissue from post-mortem Alzheimer's disease and non-disease aged matched individuals was obtained from the Victorian Brain Bank (Ethics Approval for patient tissue banking and consent: University of Melbourne HREC Approval No.: 1545740; approval for transcriptomic analysis using banked tissue Monash University MUHREC 2016-0554; patient demographics in Supplementary Table 1). Nuclei isolation was carried out using the Nuclei Isolation Kit: Nuclei EZ Prep (Sigma, #NUC101) as described in 9. Briefly, tissue samples were homogenized using a glass dounce grinder in 2 ml of ice-cold EZ PREP and incubated on ice for 5 min. Centrifuged nuclei (500 × g, 5 min and 4 °C) were washed in ice-cold EZ PREP buffer, and Nuclei Suspension Buffer (NSB; consisting of 1× PBS, 1% (w/v) BSA and 0.2 U/μl RNase inhibitor (Clontech, #2313A). Isolated nuclei were resuspended in NSB to 106 nuclei per 400 μl), filtered through a 40 μm cell strainer and counted with Trypan blue.  Nuclei enriched in Nuclei Suspension Buffer were stained with DAPI (1:1000) for nuclei isolation using the Influx cell sorter (BD Biosciences, Franklin Lakes, NJ; 70 μm nozzle, 21-22 psi). Nuclei were defined as DAPI+ singlets. Sorted nuclei were counted twice prior to loading onto the 10X Chromium (10X Genomics). Library construction was performed using the Chromium Single Cell 3′ Library & Gel Bead Kit v2 (10X Genomics, #PN-120237) with 18 cDNA pre-amplification cycles and sequencing on one high-output lane of the NextSeq 500 (Illumina). Our sequencing saturation was high, ranging from 83.2% to 97.4%. 10x Genomics Single Nuclei RNA Sequencing
GEO Accession
GSM4120427