To analyze the sequence diversity of the parental and evolved AAV libraries, DNase I-resistant viral genomes were isolated from media and amplified by Q5 Hot Start Polymerase (NEB, M0493S) for 10 to 18 cycles using the primers 5′-CCCTACACGACGCTCTTCCGATCTNNNNNGTACCTGTACTACTTGTCTCG-3′ and 5′-GACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNNAGACCATACCGGGTAAG-3′ and then sequenced with the MiSeq platform as previously described (47 (link)).
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