Preparation and titration of CsCl-banded adenovirus stocks.
Adenovirus research often requires purified high-titer virus stocks and accurate virus titers for use in experiments. Accurate titers are important for quantitative, interpretable, and reproducible results. This is especially true when there are comparisons of different mutant viruses following infection. This chapter details the large-scale preparation of adenovirus (either replication-competent or replication-defective) in spinner cultures (e.g., KB, HeLa, or 293 cells). Protocols for harvesting cells and isolation of adenovirus by CsCl banding are presented. Methods for titering adenovirus by plaque assay are presented along with a discussion of how plaque assays can be used to determine the kinetics of cell killing and cytolysis by adenoviruses.
Tollefson
Ann E
AE
Department of Molecular Microbiology and Immunology, Saint Louis University School of Medicine, St. Louis, MO, USA.
Kuppuswamy
Mohan
M
Shashkova
Elena V
EV
Doronin
Konstantin
K
Wold
William S M
WS
D000256:Adenoviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D002460:Cell Line; D002499:Centrifugation, Density Gradient / Q000379:methods; D002586:Cesium; D002712:Chlorides; D006367:HeLa Cells; D006801:Humans; D007202:Indicators and Reagents; D007624:KB Cells; D007668:Kidney
D002712:Chlorides; D007202:Indicators and Reagents; D002586:Cesium; C028019:cesium chloride