template to generate the probe from the primer the resulting 671 base product was gel purified using the qiagen ii gel extraction kit total rna 10 u03bcg was incubated with 100 ng of 32 p 671 base probe plus various levels of s1 endonuclease for 10 minutes the digested products were then analyzed on an 8 m urea 7 page gel demonstration that large aav genomes are able to effectively replicate their dna to define the packaging capacity of the aav virion the large essentially wildtype aav genomes with the inserts described in example 1 were used as described in example 1 the ligated fragments ranged in size from 600 to 1000 base pairs resulting in aav vectors ins96 0 6 ins96 0 7 ins96 0 8 ins96 0 9 and ins96 1 0 the pst i map of the cloned aav genome is shown in fig1 and an agarose gel analysis of these plasmids analyzed by pst i digestion is shown in fig2 as shown in fig1 the smallest pst i fragment u2248 0 45 kb encompasses the right aav terminal region of aav with the bglii restriction site at nucleotide 4484 map unit 96 this 0 45 kb fragment increased in size in constructs ins96 0 5 to 1 0 corresponding to the size of the 600 1000 bp fragment which was ligated into it see fig2 to observe if these altered large aav genomes were able to replicate the plasmids were analyzed by southern blot analysis both hela and sw13 cells were.