GSM6293898: NF1_BYL_1; Homo sapiens; RNA-Seq
RNA was extracted with a RNeasy Plus Mini kit (Qiagen #74136). RNA quality was analyzed on an Agilent 2100 Bioanalyzer (Agilent Technologies) using the Agilent RNA 6000 Nano Chip (Agilent #5067-1511) - RIN (RNA Integrity Number) was 8.6-10 (average 9.7). RNA was quantified by Fluorometry using the QuantiFluor RNA System (Promega #E3310). Library preparation was performed, starting from 200 ng total RNA. The library was prepared with the Illumina TruSeq stranded mRNA-seq preparation kit (Illumina #20020595) and the TruSeq RNA CD Index Plate (Illumina #20019792) according to the manufacturer's protocol. Size and quality of the library was evaluated on the Fragment Analyzer (Advanced Analytical) with the Standard Sensitivity NGS Fragment Analysis Kit (Advanced Analytical, #DNF-473). Samples were pooled to equimolar concentrations. Libraries complemented with 1% PhiX were sequenced Single-reads 76 bases (in addition: 8 bases for index 1 and 8 bases for index 2) using 2 NextSeq 500 High Output Kit 75-cycles (Illumina, Cat# FC-404-1005).