title: GSM7075623: M2 macrophages replicate_3; Homo sapiens; RNA-Seq
library_construction_protocol: RNA extraction was performed using the Qiagen Rnaeasy mini kit and RNA quality was tested using the Agilent RNA 6000 Nano kit and the Agilent 2100 Bioanalyzer. Libraries were prepared with NEBNext Ultra II Directional RNA Library Prep Kit for Illumina protocol according supplier recommendations. Briefly the key stages of this protocol are successively, the purification of PolyA containing mRNA molecules using poly-T oligo attached magnetic beads from 100ng total RNA (with the Magnetic mRNA Isolation Kit from NEB), a fragmentation using divalent cations under elevated temperature to obtain approximately 300bp fragments, double strand cDNA synthesis and finally Illumina adapters ligation and cDNA library amplification by PCR for sequencing. Sequencing was carried out on Paired End 100b reads on Illumina NovaSeq6000. Image analysis and base calling was performed using Illumina Real Time Analysis with default parameters