RNA-seq; Homo sapiens; Umbilical cord blood
cDNA was synthesized from the total cellular RNA using Clontech SmartSeq v4 reagents from 1ng of RNA. Full length cDNA was fragmented to a mean size of 150bp with a Covaris M220 ultrasonicator and Illumina libraries were prepared from 2ng of sheared cDNA using Rubicon Genomics Thruplex DNAseq reagents according to manufacturers protocol. The finished dsDNA libraries were quantified by Qubit fluorometer, Agilent TapeStation 2200, and RT-qPCR using the Kapa Biosystems library quantification kit. Uniquely indexed libraries were pooled in equimolar ratios and sequenced on an Illumina NextSeq500 run with single-end 75bp reads at the Dana-Farber Cancer Institute Molecular Biology Core Facilities.