GSM7184299: Ly8_het_A9_2; Homo sapiens; RNA-Seq
RNA was isolated using Direct-zolTM RNA MicroPrep kit (Zymo Research) and each sample was normalized to 10 ng/µl. A total of 10 ng of RNA per sample were subjected to RNA sequencing using a version of the prime-seq protocol (Janjic et. al. BioRXiv, 2021). A full step-by-step protocol can be found on protocols.io (https://www.protocols.io/view/prime-seq-s9veh66). Prime-seq is a three prime counting method that introduces a sample specific barcode sequence and unique molecular identifiers (UMI) for accurate quantification of gene expression. Illumina paired end sequencing was performed on an HiSeq 1500 instrument, where the first read was 16 bases long and covered the sample barcode and UMI, and the second read was 50 bases long and was used to identify the gene.