A rapid method for screening vaccinia virus recombinants.
A rapid and small-scale method for screening vaccinia virus recombinants employing micrococcal nuclease is described. This protocol utilizes the differential sensitivity of cellular and viral DNA to the nuclease, which can be selectively activated by addition of Ca2+ and inactivated by elimination of Ca2+. Two to five micrograms of viral DNA can be obtained from one infected L cell plate (50 mm) after overnight incubation.
Division of Cell Biology, John Curtin School of Medical Research, Australia.
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Lai
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Chu
D000818:Animals; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D066298:In Vitro Techniques; D007739:L Cells / Q000648:ultrastructure; D051379:Mice; D012150:Polymorphism, Restriction Fragment Length; D014616:Vaccinia virus / Q000235:genetics*
D004274:DNA, Recombinant; D004279:DNA, Viral