title: LAM-PCR to profile vector genome integration of embryo explants treated with rAAV-CRISPR-Cas9
design: Whole-genomic DNAs were extracted from snap-frozen and powdered tissues from experimental E16.5 embryos.  Genomic material (20 ug total input) was fragmented by Taq I digestion. Fragmented DNAs were subjected to phenol-chloroform extraction and ethanol precipitation to purify the fragmented material.  Template DNAs were next subjected to 80 cycles of LAM-PCR.  Single-strand, biotinylated PCR products were next captured on magnetic beads, enriched, and ligated to a bridge adapter by on-bead ligation.  Nested PCR (30-cycles) to generate SMRT-sequencing libraries was next carried out using asymmetrically barcoded. Individual libraries were then subjected to 0.6X AMPurePB bead purification, pooled, and submitted for standard SMRT-sequencing analysis