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Analysis of replicative intermediates of adeno-associated virus through hirt extraction and southern blotting.

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).
Publication status:
Published
Peer review status:
Peer reviewed
Version:
Publisher's version

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Publisher copy:
10.21769/BioProtoc.2271

Authors


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Department:
Oxford,MSD,,NDMJenner Institute
Zarate-Perez, F More by this author
Agundez, L More by this author
Jolinon, N More by this author
Michael Linden, R More by this author
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Pfizer Rare Diseases Consortium More from this funder
Publisher:
Bio-Protocol Publisher's website
Journal:
Bio-Protocol Journal website
Volume:
7
Issue:
9
Publication date:
2017-05-05
DOI:
ISSN:
2331-8325
Pubs id:
pubs:707852
URN:
uri:72bc62c2-af70-43bf-ad9d-acef9511f3a0
UUID:
uuid:72bc62c2-af70-43bf-ad9d-acef9511f3a0
Local pid:
pubs:707852

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