title: GSM4314510: GOS_rep2; Homo sapiens; RNA-Seq
library_construction_protocol: Cells were gently washed in situ twice with ice cold sterile PBS followed by addition of 500 μl of TRIzol™ reagent (ThermoFisher Scientific, Paisley, UK) for harvesting by directly scraping the cells from the culture surface. Harvested cells were treated for total RNA extraction according to TRIzol™ reagent manufacturer's protocol. Purified RNAs were quality-assessed using Agilent TapeStation 2200 and  were subsequently aliquoted to limit freeze-thaw cycles and stored in −80°C.  Library was prepared according to Illumina protocol. Briefly, mRNA was purified from 1 μg of total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module (E7490; New England Biolabs, Ipswich, UK). Indexed sequencing libraries were prepared using the NEBNext Ultra directional RNA Library Preparation Kit for Illumina (E7760; New England Biolabs, Ipswich, UK) and NEBNext Multiplex Oligos for Illumina, Index Primers Sets 1 and 2 (E7335 and E7500; New England Biolabs, Ipswich, UK). Libraries were quantified using the Qubit Fluorometer and the Qubit dsDNA HS Kit (Q32854; ThermoFisher Scientific, Paisley, UK). Library fragment-length distributions were analysed using the Agilent TapeStation 4200 and the Agilent High Sensitivity D1000 ScreenTape Assay (5067-5584 and 5067-5585; Agilent, Stockport, UK). For sequencing, libraries were pooled in equimolar amounts and final library quantification performed using the KAPA Library Quantification Kit for Illumina.
attributes.tag: GEO Accession
attributes.value: GSM4314510