- Analysis of Replicative Intermediates of Adeno-associated Virus through Hirt Extraction and Southern Blotting.
Analysis of Replicative Intermediates of Adeno-associated Virus through Hirt Extraction and Southern Blotting.
Bio-protocol (2017-07-01)
Martino Bardelli, Francisco Zarate-Perez, Leticia Agundez, Nelly Jolinon, R Michael Linden, Carlos R Escalante, Els Henckaerts
PMID28660237
ABSTRACT
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).
MATERIALI
N° Catalogo
Marchio
Descrizione del prodotto
Sigma-Aldrich
Sodio dodecil solfato, BioReagent, suitable for electrophoresis, for molecular biology, ≥98.5% (GC)
Sigma-Aldrich
Miscela di fenolo-cloroformio-alcol isoamilico, BioUltra, for molecular biology, 25:24:1