GSM7405310: sample_2219; Homo sapiens; RNA-Seq
RNA was extracted using TRIzol reagent (Thermo Fisher) following the standard protocol. Samples with acceptable RNA quality (RIN > 7; assessed with Agilent 6000 RNA Nano chip) and quantity (200ng or more; assessed with Invitrogen Qubit High Sensitivity RNA assay) were used in library preparation. NEBNext® Ultra™ II Directional (stranded) RNA Library Prep Kit for Illumina (NEB #E7760L) was used as the core kit and coding genes were enriched using the NEBNext Poly (A) mRNA Magnetic Isolation Module (NEB #E7490). RNA fragmentation time: 10 min. PCR cycles: 12. NEBNext Multiplex Oligos for Illumina Unique Dual Index Primer Pairs (NEB #6446/L) were used to index each sample for multiplexing purposes. Final libraries were quantified using Qubit High Sensitivity DNA assay and pooled to yield ~20 million clusters/sample. mRNA-seq libraries were sequenced using Illumina NovaSeq 6000 flow cell with Paired-End 100 bp format to ~20 million clusters/sample.