In response to the NHLBI RFP for Transcriptome RNAseq of a minimum of 1,000 samples, we are proposing to profile these samples using the Illumina TruSeq Stranded HT mRNA kit for library preparation, and to sequence the libraries on the Illumina NovaSeq using paired-end 2x100bp reads with eight-base unique dual index barcodes at a minimum depth of 75M mapped reads per blood derived samples and 50M mapped reads per tissue derived or globin-depleted blood derived samples, delivered as CRAM, with a goal of 17,100 samples total. The details of this process are described below, together with data supporting our approach.