title: Old Donor #2
design: Library preparation was performed according to Illuminas TruSeq Nano DNA sample preparation protocol. The samples were sheared on a Covaris S220 to ~450bp, following the manufacturers recommendation, and uniquely tagged with one of Illuminas TruSeq LT DNA barcodes to enable library pooling for sequencing. The finished libraries were quantitated using Invitrogens Picogreen assay and the average library size was determined on a Bioanalyzer 2100, using a DNA 7500 chip (Agilent). Library concentrations were then normalized to 4nM and validated by qPCR on a ViiA-7 real-time thermocycler (Applied Biosystems), using qPCR primers recommended in Illuminas qPCR protocol, and Illuminas PhiX control library as standard. The libraries were then pooled at equimolar concentrations and sequenced in one lane on an Illumina HiSeq2500 sequencer in rapid mode at a read-length of 250bp paired-end.