title: GSM7675749: PBMC, Tconv, EPF/infertile, subject2; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was isolated using the Qiagen RNeasy Micro Plus Kit according to the manufacturer's protocols. 46-548ng (mean 200ng) of total RNA was subjected to cDNA library synthesis using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, Ipswich, MA), NEBNext Ultra II Directional RNA Library Prep Kit from Illumina (San Diego, CA) and NEBNext Multiplex Oligos for Illumina, according to the manufacturer's protocols. The libraries were quantified with the Universal KAPA Library Quantification Kit (Roche, Basel, Switzerland) and those with sufficient DNA (≥ 2 nM) were pooled to equimolar concentration. Paired-end 150bp sequencing was performed at GENEWIZ (Azenta Life Sciences, Chelmsford, MA) on an Illumina Hi-Seq platform across 5 lanes. Libraries were sequenced to an average read depth of 56.3 million reads per sample.