Analysis of Replicative Intermediates of Adeno-associated Virus through Hirt Extraction and Southern Blotting

Autor: Martino Bardelli, Francisco Zarate-Perez, Leticia Agundez, Nelly Jolinon, R. Linden, Carlos Escalante, Els Henckaerts
Jazyk: angličtina
Rok vydání: 2017
Předmět:
Zdroj: Bio-Protocol, Vol 7, Iss 9 (2017)
Druh dokumentu: article
ISSN: 2331-8325
DOI: 10.21769/BioProtoc.2271
Popis: Adeno-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).
Databáze: Directory of Open Access Journals