Retroviral insertions amplified by LM-PCR and sequenced on 454 FLX
Genomic DNA from human hematopoietic progenitors infected with MLV retroviral vectors was extracted and digested with MseI. 3’ LTR vector-genome junctions were amplified by linker-mediated PCR (LM-PCR), adapted to the GS-FLX Genome Sequencer pyrosequencing platform. Pooled LM-PCR amplicons were quantified, size-fractionated by SPRI beads, and sequenced according to the GS-FLX manufacturer's instructions.