GSM4025081: eSF_Mock_1; Homo sapiens; RNA-Seq
Total RNA was extracted from eSF or eEC cultures (1 μg of total RNA from each sample) using the High Pure RNA Isolation kit (Roche) according to the manufacturer's protocol. Quality of isolated RNA was confirmed using Nanodrop for RNA purity (OD 260/280) and Bioanalyzer (Agilent) for RNA integrity. RNA samples with an RNA Integrity Number (RIN) > 6 were used for library preparation for mRNA sequencing. All RNA sequencing (RNAseq) procedures were performed by Novogene. Library preparation was performed using 1 μg of total RNA from each sample with the NEBNext Ultra RNA Library Prep Kit (New England BioLabs, Inc.). Briefly, polyadenylated (polyA) RNAs were selected using NEBNext Magnetic Oligo d(T)25 Beads (New England BioLabs, Inc.). Following polyA selection, RNA was fragmented and primed for first-strand cDNA synthesis, followed by second-strand synthesis. Double stranded cDNA was purified using Agencourt AMPure XP Beads (Beckman Coulter), the fragments were end-repaired, 3' adenylated, and NEBNext Adaptor with hairpin loop structure (Illumina) was ligated to prepare for hybridization. cDNA fragments ~350 bp in length were selected using the Agencourt AMPure XP system (Beckman Coulter) and PCR amplification was performed to create the final cDNA library. Paired-end RNAseq, at >20 million reads per sample was sequenced on an Illumina HiSeq platform (Illumina), per the manufacturer's instructions.
GEO Accession
GSM4025081