Analysis of Replicative Intermediates of Adeno-associated Virus through Hirt Extraction and Southern Blotting
2017; American Academy of Arts and Sciences; Volume: 7; Issue: 9 Linguagem: Inglês
10.21769/bioprotoc.2271
ISSN2331-8325
AutoresMartino Bardelli, Francisco Zárate-Pérez, Leticia Agúndez, Nelly Jolinon, R. Michael Linden, Carlos Escalante, Els Henckaerts,
Tópico(s)CRISPR and Genetic Engineering
ResumoAdeno-associated virus (AAV) is a small single-stranded DNA virus that requires the presence of a helper virus, such as adenovirus or herpes virus, to efficiently replicate its genome. AAV DNA is replicated by a rolling-hairpin mechanism (Ward, 2006), and during replication several DNA intermediates can be detected. This detailed protocol describes how to analyze the AAV DNA intermediates formed during AAV replication using a modified Hirt extract (Hirt, 1967) procedure and Southern blotting (Southern, 1975).
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