GSM6617476: HD98_Empty vector; Homo sapiens; RNA-Seq
RNA was prepared from 500,000 cells with the RNeasy Plus Kits (Qiagen) including a DNAse treatment step, according to the manufacturer's instructions. RNA quality was assessed by capillary electrophoresis using High Sensitivity RNA reagents with the Fragment Analyzer (Agilent Technologies) and the RNA concentration were measured by using both spectrophometry using the Xpose (Trinean) and Fragment Analyzer capillary electrophoresis. Libraries were prepared starting from 200ng of total with the Universal Plus mRNA-Seq kit (Nugen) as recommended by the manufacturer (libraries can be prepared from 10 ng to 1µg of starting total RNA). The oriented cDNA produced from the poly-A+ fraction were sequenced on a NovaSeq6000 from Illumina (Paired-End reads 100 bases + 100 bases). A total of ~50 millions of passing filter paired-end reads was produced per library.