GSM8265715: Monocytes, control plasmid (pcDNA), rep4; Homo sapiens; RNA-Seq
Monocytes were harvested by centrifugation and washed with ice-cold PBS. RNA was purified from cells using the miRNeasy kit (Qiagen, #217004) according to manufacturer's instructions with additional DNase (Qiagen, #79254) digestion. Quality control and quantification of the isolated RNA were performed using Agilent fragment analyzer DNF-471 Standard Sensitivity RNA Kit. From each sample, 100 ng was used as input for library preparation using the Illumina Truseq totalRNA stranded Kit (#20020597). Ready prepped library was monitored again using the fragment analyzer and the DNF-473 Standard Sensitivity NGS Fragment Kit. Based on the measured concentration, libraries were then diluted to 5 nmol/l and equimolar amounts of each sample were plexed at 1.25 nmol/l. Sequencing was performed as paired-end 2 x 150 bp (S4-Flowcell) on a Novaseq 6000 (Illumina).