ISOLATION OF GENOMIC SEQUENCES FLANKING A RETROVIRAL INSERTION SITE USING A NOVEL PCR-BASED METHOD

A novel PCR approach for the isolation of genomic sequences that flank retroviral insertion sites uses two standard primers and defined PCR conditions. The method is based on the directional cloning of digested genomic DNA into the multiple cloning site of pUC-based plasmids, iso lation of recombinant plasmid DNA from pooled colonies and PCR amplifi cation using primers specific for the pUC vector and retroviral LTR se quences. The method is simple and can be adapted to enable rapid isola tion of any genomic sequences tagged by insertion of foreign DNA.