Figure 1.

Illumina MiSeq MGS-PCR. Genomic DNA is sheared, resulting in smaller genomic fragments that may contain the vector provirus. Fragments are blunt ended and linker cassettes ligated to the ends of the fragment. Thirty rounds of exponential PCR are conducted with a biotin-tagged LTR primer and linker-specific primer. The resulting amplicons undergo selection of biotin-tagged amplicons with streptavidin beads to enrich for fragments containing LTRs. Amplicons are then recovered and 30 rounds of nested PCR performed using an Illumina universal forward LTR-specific primer and an Illumina universal reverse linker-specific primer are performed. The resulting amplicons are gel isolated and ready for sequencing on the MiSeq. Color images available online at www.liebertpub.com/hgtb