GSM8191638: 12_month_old_WT3; Mus musculus; RNA-Seq
RNA isolation was carried out from 40 cryosections taken from the tongue median region, N=4 wild type and N=4 A17/+ female mice. Cryosections were collected in a MagNA lyser green beads tubes (Roche, 3358941001), and tissue was homogenized in QIAzol lysis reagent (Qiagen, 79306) using the MagNA 406 Lyser. Subsequently, total RNA was isolated with the miRNeasy Mini Kit (217004, Qiagen) using the manufacturer's protocol. DNA was removed using RNAse-free DNAse set (Qiagen, 79254) using the manufacturer's protocol. RNA integrity was assessed with the Agilent 2100 Bioanalyzer using Eukaryote Total RNA Nano chips according to the manufacturer's protocol (Agilent 415 BioAnalyzer, 824.070.709) The generation of cDNA for the RNA-seq data was carried out with the mcSCRB-seq protocol, with the following modifications to the workflow: instead of single cells, we used 1–100ng isolated total RNA, 7.5% PEG8000 was replaced with 0.5% polyvinylpyrrolidone, and the oligo-dT primer was replaced with the SCIFI_LIG384_001 primer. All samples were barcoded during reverse transcription using a well barcode. Illumina paired-end sequencing of the library was done on a NovaSeq 6000 using paired-end 150-bp sequencing and v1.5 chemistry. After sequencing, read 1 will contain the cDNA information and read 2 will contain only the unique molecular identifier