Insertion sites analysis for CART therapy

To assess whether the different methods of cell preparation affected distributions of vector integration sites, samples were analyzed by ligation-mediated PCR. Experimental infections were carried out in resting and activated cells, and DNA purified. DNAs were sheared by sonication, DNA adaptors ligated onto the free DNA ends, and PCR carried out between the adaptor and the edge of the integrated vector. PCR products were then subjected to sequencing, DNA sequences were mapped back onto the human genome, and vector integration site distributions compared.