Efficient identification of recombinant adenoviruses by direct plaque screening.
With the increasing use of adenoviral vectors for gene transfer and gene therapy, it is crucial to produce specific recombinant adenoviruses more efficiently. One of the most time-consuming steps is to screen the unique recombinant adenovirus among the plaques, in which each plaque has to be amplified individually in kidney 293 cells in order to obtain enough adenoviruses for DNA extraction and subsequent identification. We have developed a fast and simple way to screen recombinant adenoviruses by direct plaque screening. The direct plaque-screening method employed DNA obtained from the viral plaque itself for PCR amplification and subsequent adenoviral recombinant identification. The time and labor involved in these steps has been significantly reduced.
Lu
Y
Y
Department of Urology, University of Tennessee, Memphis 38163, USA.
Zhang
Y
Y
Steiner
M S
MS
D000256:Adenoviridae / Q000737:chemistry / Q000235:genetics*; D019677:Artificial Gene Fusion / Q000379:methods; D004274:DNA, Recombinant / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D005822:Genetic Vectors / Q000032:analysis / Q000235:genetics*; D016133:Polymerase Chain Reaction
D004274:DNA, Recombinant; D004279:DNA, Viral