GSM4387148: Extracellular Vesicles_treated_Replicate_1; Bos taurus; miRNA-Seq
Total RNA enriched with miRNAs was isolated from the purified EVs using Norgen exosomes RNA kit (Norgen, Canada) according to the manufacturer´s instruction. The library preparation was done using the QIAseq miRNA Library Kit (QIAGEN). A total of 100ng total RNA was converted into miRNA NGS libraries. Adapters containing UMIs were ligated to the RNA. Then RNA was converted to cDNA. The cDNA was amplified using PCR (16 cycles) and during the PCR indices were added. After PCR the samples were purified. Library preparation QC was performed using either Bioanalyzer 2100 (Agilent) or TapeStation 4200 (Agilent). Based on quality of the inserts and the concentration measurements the libraries were pooled in equimolar ratios. The library pool(s) were quantified using qPCR. The library pool were then sequenced on a NextSeq550 sequencing instrument according to the manufacturer instructions. Raw data was de-multiplexed and FASTQ files for each sample were generated using the bcl2fastq software (Illumina inc.). FASTQ data were checked using the FastQC tool.
GEO Accession
GSM4387148