SMRT-Tag preparation of Homo sapiens: 40ng HG003 NIST reference material
40ng Genomic DNA is tagmented using 5pmol of custom-loaded Tn5 transposase with hairpin adapters compatible with PacBio SMRT sequencing in 10mM TAPS-NaOH pH 7.5, 5mM MgCl2, and 10% DMF at 37C for 30 minutes. Fragments are cleaned up using a 2X SPRI and gap-repaired using a polymerase / ligase combination (Phusion polymerase / Taq ligase) at 37C for 1hr. Libraries are then digested with exonuclease III at 37C for 1hr to remove un-repaired, non-productive templates.