GSM5256869: v1068_12: vol1068_third_inf_C6; Homo sapiens; RNA-Seq
Total whole blood RNA was extracted using the Tempus™ Spin RNA isolation reagent kit (ThermoFisher #4380204) according to manufacturer's instructions. RNA integrity assessed using an Agilent Bioanalyzer 2100 (RNA 6000 nano kit Agilent #5067-1511). Our average RNA integrity number (RIN) was 9 (98% of samples > 8, lowest RIN 7). Libraries were constructed by Edinburgh Genomics (Edinburgh, UK) from approximately 500 ng of RNA using the TruSeq® Stranded mRNA library prep kit (Illumina #20020595). Stranded library preparation allows transcripts expression to be estimated more accurately; in particular it is more effective in quantifying anti-sense gene expression, properly assigning transcripts to putative coding genes and to resolve ambiguity in reads from overlapping genes (Zhao, S., et al., Comparison of stranded and non-stranded RNA-seq transcriptome profiling and investigation of gene overlap. BMC Genomics, 2015, PMID: 26334759). Libraries were sequenced using the NovaSeq 6,000 Illumina® platform yielding 50 bp paired end (PE) reads.