GSM2886340: Leicester_with_progressor_longitudinal_Sample74; Homo sapiens; RNA-Seq
Total RNA was isolated from 1 ml whole blood using the MagMAX™ for Stabilized Blood Tubes RNA Isolation Kit (Applied Biosystems/Thermo Fisher Scientific) according to the manufacturer's instructions. Globin RNA was depleted from total RNA (1.5–2 µg) using the human GLOBINclear kit (Thermo Fisher Scientific) according to manufacturer's instructions. RNA yield of total and globin-reduced RNA was assessed using a NanoDrop™ 8000 spectrophotometer (Thermo Fisher Scientific). Quality and integrity of total and globin-reduced RNA were assessed with the HT RNA Assay reagent kit (Perkin Elmer) using a LabChip GX bioanalyser (Caliper Life Sciences/Perkin Elmer) and assigned an RNA Quality Score (RQS). Samples (200 ng) with an RQS > 6 were used to prepare a cDNA library using the TruSeq Stranded mRNA HT Library Preparation Kit (Illumina). The tagged libraries were sized and quantitated in duplicate (Agilent TapeStation system), using D1000 ScreenTape and reagents (Agilent), normalised, pooled and then clustered using the HiSeq® 3000/4000 PE Cluster Kit (Illumina). The libraries were imaged and sequenced on an Illumina HiSeq 4000 sequencer using the HiSeq® 3000/4000 SBS kit (Illumina) at a minimum of 25 million paired end reads (75 bp) per sample.
GEO Accession
GSM2886340