title: Sequencing of P0
design: Genomic DNA was purified from 2mls of whole blood from P0 using the  DNeasy Kit.  DNA fragments of approximately 30kb were sheared using a Covaris S2 acoustic sonicator using conditions recommended by Helicos.  The intensity and time settings were varied to maximize the fragment yield in the 100-500bp range.  Sheared DNA fragments were further processed using a Microcon 30 (Millipore) column to remove small fragments and treated with T4 polynucleotide kinase to maximize 3’-OH groups at the ends of the fragments.    Terminal transferase tailing with dATP, 3’-terminal blocking, concentration and quantitation used protocols provided by Helicos.  Approximately 200 pmoles of dA-tailed molecules were loaded onto each lane of two flowcells.  This represents approximately 25pg of DNA per lane.  Control oligonucleotides provided by Helicos were loaded into one lane on each flowcell for sequencing run quality control.   Minor modifications to the DNA sample preparation using size selected fragments of 100-300bp or using Dynal (Invitrogen) oligo-dT magnetic beads to enrich for polydA-containing molecules did not improve the sequence output.