Integration site isolation of LV-w1.6 WASp gene therapy vector from adult homo sapien WAS patient
DNA was randomly sheared via ultrasonication and DNA linkers were ligated to the cleaved ends. DNA fragments were then amplified by ligation-mediated PCR, using primers annealing to the LTR of the vector and to the linker to isolate and enrich for host-vector junctions. PCR was performed in quadruplicate for each sample. Amplicons were sequenced using an Illumina MiSeq instrument. Sequence reads were aligned to the human genome (hg18) to identify vector integration sites.