GSM8286282: Scr1 scRNAseq; Mus musculus; RNA-Seq
FACS-sorted cells were processed using the Smart-seq2 protocol. Briefly, RNA was reverse-transcribed using biotinylated oligo-dT primers (IDT DNA, Belgium) and biotinylated template-switching (TSO) oligonucleotides (Exiqon, QIAGEN, Denmark) and subsequently cDNA was pre-amplified using the KAPA HiFi Hot Start DNA Polymerase (KAPA Biosystems, Roche Diagnostics, Belgium) and biotinylated ISPCR primers (IDT DNA, Belgium). cDNA was purified using AMPURE XP Agencourt beads (Beckman Coulter, France), concentration was calculated with the Quantifluor dsDNA kit (Promega, Netherlands) and adjusted on an Echo 525 Liquid Handler (Labcyte, CA, USA). 100 pg cDNA per cell were used for library preparation with Nextera XT DNA Library Preparation Kit (Illumina, CA, USA). Following purification on magnetic beads the libraries were multiplexed and sequenced using the Illumina NextSeq500 platform (Illumina, CA, USA).