GSM8136760: Control-2; Homo sapiens; RNA-Seq
Total RNA extraction was performed using AllPrep DNA/RNA Mini Kit (QIAGEN), following the manufacturer's protocol. RNA quality was assessed using an Agilent 5200 Fragment Analyzer, using the Agilent HS RNA (15NT) kit. The NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) was used to isolate intact poly(A)+ RNA from 1µg of previously isolated total RNA. Libraries were prepared using the Ultra II Directional RNA Library Prep Kit for Illumina and Unique Dual Index Primer Pairs were applied following the vendor protocols from New England Biolabs. Samples were sequenced in a single run on an Illumina NextSeq 550 using the NextSeq 500/550 High Output Kit v2.5 (300 Cycles) to produce 150–base pair-end reads with a mean sequencing depth of at least 60 million reads per sample.