GSM8988816: T126_B_MIN; Homo sapiens; RNA-Seq
Cell lysates were first homogenized using QIAshredder columns followed by RNA isolation using the Qiagen RNeasy Plus kit according to manufacturers instructions, including on-column DNase treatment. Poly(A)-enriched mRNA libraries were prepared as follows. 300ng per sample was processed according to NEBNext Ultra II Directional RNA Library Prep sample preparation protocol (Part # E7760L) with the NEBNext Poly(A) mRNA Magnetic Isolation Module (Part # E7490). 1µL top stock of each purified final library was run on an Agilent TapeStation HS D1000 tape (Agilent). The libraries were quantified using Quant-iT dsDNA HS Assay Kit (ThermoFisher Scientific) and were pooled at equimolar ratios after size adjustment. The final pool was run on an Agilent Bioanalyzer dsDNA High Sensitivity chip and quantified using NEBNext Library Quant Kit for Illumina (NEB). The quantified pool was hybridized at a final concentration of 2.75pM and sequenced paired-end 2x50 bp on the Illumina NovaSeq6000 platform using an S2 flowcell.