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Updated: Feb 27, 2026

Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors
Published on: April 5, 2024
Analysis of Replicative Intermediates of Adeno-associated Virus through Hirt Extraction and Southern Blotting
Martino Bardelli1, Francisco Zarate-Perez2, Leticia Agundez1
1Department of Infectious Diseases, King's College London, London, United Kingdom.
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).
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