GSM4906537: Donor r B. theta dCPS LPS rep2; Homo sapiens; RNA-Seq
Total RNA was extrated by adding a 3x volume of TRIzol™ LS reagent (Ambion) or TRI Reagent® (Zymo Research) to the cells. The suspension was mixed by pipetting to lyse the cells, followed by RNA isolation using the Direct-zol™ RNA Miniprep kit (Zymo Research) including an on-column DNase I digestion step. RNA-seq libraries were prepared from purified RNA using the Stranded mRNA Prep Ligation kit (Illumina) in 96-well format, following the manufacturer's instructions. Input RNA amounts were held constant for all samples for a given donor, between 300 and 600 ng per reaction. Final libraries were validated and quantified using the 2100 Bioanalyzer (Agilent) using the High Sensitivity DNA kit (Agilent). Paired-end 100 sequencing was performed on a HiSeq 4000 (Illumina).