RAPID SMALL-SCALE ISOLATION OF HERPES-SIMPLEX VIRUS-DNA

A method has been developed for the rapid isolation of herpes simplex virus DNA analogous to miniprep methods for bacterial plasmid isolatio n. Infected Vero cells are lysed with three freeze-thaw cycles, and th e nuclei are removed by centrifugation. DNA is released from the virio ns in the supernatant by proteinase K digestion. Then the DNA is extra cted with phenol/chloroform and precipitated with ethanol. This method requires only small amounts of infected cells as a source of viral DN A, does not use radioactivity, and routinely produces DNA of sufficien t purity to be used for restriction fragment length polymorphism (RFLP ) analysis on ethidium-stained gels.