Analysis of Replicative Intermediates of Adeno-associated …
2017年5月5日 · 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). Keywords: AAV; Adeno-associated virus; DNA replication; Replicative intermediates; Southern blot.
复制性完整型AAV的检验方法 | 派真生物 - PackGene Biotech
Southern blotting： Southern blotting是一种检验AAV样品中的基因组完整性的方法。通过将AAV样品中的DNA进行限制性内切酶消化、电泳分离、转移至膜上、杂交等步骤，可以检测AAV基因组的完整性。
High levels of AAV vector integration into CRISPR-induced DNA …
2019年9月30日 · Southern blotting of genomic DNA of U2-OS cells revealed integration events of various sizes after AAV-λ465 transduction, including shorter and longer integrants than the size of the AAV-λ465 ...
Formation of AAV Single Stranded DNA Genome from a Circular
2011年8月10日 · Molecular analysis of low molecular weight DNA by Southern blotting revealed that single stranded DNA is formed and that the plasmid is entirely replicated. The ssDNA contains the ITRs, URA3 gene and also vector sequences suggesting the …
Southern blot analysis of adeno-associated virus (AAV) genome ...
Adeno-associated virus (AAV) vectors offer unique advantages, such as tissue tropism, specificity in transduction, elicited a relatively low immune responses, no incorporation into...
Analysis of Replicative Intermediates of Adeno-associated Virus …
2017年5月5日 · AAV DNA is replicated by a rolling-hairpin mechanism , 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 procedure and Southern blotting (Southern, 1975).
Characterization of Genome Integrity for Oversized Recombinant AAV ...
2009年11月10日 · A plasmid replication origin and an ampicillin-resistant marker were included in the vector to facilitate the recovery of circularized, post-transduction AAV genome. Southern dot-blot analysis and silver staining confirmed that rAAVs could be produced at varying vector size.
Southern blot analysis of rescue and replication of the AAV and …
Southern blot analysis of rescue and replication of the AAV and the vector DNA sequences from recombinant plasmids containing ITR deletions. DNA-mediated transfections were carried...
A novel class of self-complementary AAV vectors with multiple ...
2024年3月14日 · Self-complementary AAV vectors (scAAV) use a mutant inverted terminal repeat (mITR) for efficient packaging of complementary stranded DNA, enabling rapid transgene expression. However, inefficient resolution at the mITR leads to the packaging of monomeric or subgenomic AAV genomes.
Characterization of genome integrity for oversized recombinant AAV ...
A plasmid replication origin and an ampicillin-resistant marker were included in the vector to facilitate the recovery of circularized, post-transduction AAV genome. Southern dot-blot analysis and silver staining confirmed that rAAVs could be produced at varying vector size.