GSM3356468: KM_1; Homo sapiens; RNA-Seq
200 μl of non-hemolyzed cord blood plasma was used to isolate RNA, using the miRCURY RNA isolation kit (Exiqon) according to the manufacturer's protocol. RNA was eluted in 25 μl. A standard overnight RNA ethanol precipitation step was then performed and RNA was re-suspended in 10 μl RNase free water to concentrate. RNA integrity and quantity was determined using the small RNA detection kit (DNF-470) on a Fragment Analyzer (Advanced Analytical Technologies). RNA (5 μl) from each sample was used to construct small RNA libraries using the Illumina TruSeq Small RNA library kit. Due to the small amount of input RNA the protocol was modified slightly (all kit reagents were halved) to prevent extensive adapter dimer formation. Libraries were size selected using a Pippin Prep with 3% agarose dye free cassettes and size selection was validated using a 2100 High sensitivity DNA Bioanalyzer chip (Agilent). The concentration of each library was determined using the HS-dsDNA kit for Qubit, libraries were pooled and pooled libraries were sequenced at the Trinseq Facility at the Institute for Molecular Medicine at St. James Hospital Dublin on an Illumina miSeq.