Human genome Brazilian individual
Twenty micrograms of genomic DNA purified from blood sample were sheared using HydroShear to generate fragments with an average size of 2.0 kb. DNA fragments were then processed to generate blunt ends and ligated to specific adaptors. DNA fragments were size-selected in agarose gels and subsequently circularized by ligation of a biotinylated internal adaptor. After removing non-circularized fragments, circularized DNA was treated with DNA polymerase I for nick-translation, followed by digestion with T7 exonuclease and S1 nuclease, which generated tags longer than 50 bp from the adaptor edges. Digested products were ligated with P1 and P2 adaptors, purified and amplified using a PCR protocol with 12 cycles. A total of 96 picograms of the resulting library was then used for emulsion PCR. Approximately 300 million beads from each library were deposited on one slide, followed by 50 bp mate pair sequencing on a SOLiD3 instrument, according to the manufacturer's protocol.