GSM8413148: CSMN_WT_08-05-2019; Homo sapiens; RNA-Seq
mRNA was isolated from 30 ng total RNA by poly-dT enrichment using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) according to the manufacturer's instructions. mRNA was isolated from 30 ng total RNA by poly-dT enrichment using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) according to the manufacturer's instructions. Samples were then directly subjected to the workflow for strand-specific RNA-Seq library preparation (Ultra II Directional RNA Library Prep, NEB). For ligation NEB Next Adapter for Illumina of the NEB Next Multiplex Oligos for Illumina Kit were used. After ligation, adapters were depleted by an XP bead purification (Beckman Coulter) adding the beads solution in a ratio of 0.9:1 to the samples. During the following 15 cycles PCR enrichment, unique dual index primer (Truseq design) were incorporated. After two XP bead purifications (with a ratio of 0.9:1 to the sample volume), libraries were quantified using the Fragment Analyzer (Agilent). Libraries were sequenced on an Illumina Nextseq 550 system in 75 bp single-end mode to an average depth of 30 million fragments per library.