GSM4980268: IL1RIKO_2y_1; Mus musculus; RNA-Seq
The RNA extraction was performed following the protocol of ArcturusTM PicoPureTM RNA isolation kit (Applied Biosystems). 4000-5000 cells were sorted directly in the 100ul of extraction buffer of the kit. Quality and quantity of RNA was measured using Agilent 4200 TapeStation System. The libraries were prepared following the Smart-seq2 protocol. Following reverse transcription, the quality of the cDNAs was evaluated using an Agilent 2100 Bioanalyzer. 0,5 ng of cDNA from each sample were tagmented and amplified using Illumina Nextera XT kit. The resulting libraries were pooled, double-sided size selected (0.5x followed by 0.8x ratio using Beckman Ampure XP beads) and quantified using an Agilent 4200 TapeStation System. The pool of libraries was sequenced in an Illumina NovaSeq6000 sequencer (single-end 100 bp) with a depth of around 20 Mio reads per sample. 