title: Whole genome seq of human: adult male B-Lymphocyte
design: Based on concentration measured by fluorescence, a 100ng aliquot of the genomic DNA will be prepared using the Illumina TruSeq Nano DNA HT library preparation kit with 350bp insert size.  Intact genomic DNA will be sheared using the Covaris sonicator (adaptive focused acoustics), followed by end-repair.  The size selected fragments will then have adenines added to the 3 ends of the DNA, Illumina sequence adaptors ligated onto the fragments, followed by PCR amplification, clean-up, and final library QC.  A majority of the steps in this process will be carried out using the Caliper SciClone NGSx workstation, a robotics system developed and validated for automated library preparation.  The library QC will include a measurement of the average size of library fragments using the BioAnalyzer, estimation of the total concentration of DNA by PicoGreen, and a measurement of the yield and efficiency of the adaptor ligation process with a quantitative PCR assay (Kapa) using primers specific to the adaptor sequence.  Sequencing will be performed on the HiSeq X instrument; libraries will be loaded onto the HiSeq X flowcell for clustering on the cBot using the instrument specific clustering protocol.