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 isolation. Infected Vero cells are lysed with three freeze-thaw cycles, and the nuclei are removed by centrifugation. DNA is released from the virions in the supernatant by proteinase K digestion. Then the DNA is extracted with phenol/chloroform and precipitated with ethanol. This method requires only small amounts of infected cells as a source of viral DNA, does not use radioactivity, and routinely produces DNA of sufficient purity to be used for restriction fragment length polymorphism (RFLP) analysis on ethidium-stained gels.
Kintner
R L
RL
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison 53706.
Brandt
C R
CR
D000818:Animals; D002522:Chlorocebus aethiops; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D005069:Evaluation Studies as Topic; D005615:Freezing; D012150:Polymorphism, Restriction Fragment Length; D011995:Recombination, Genetic; D018139:Simplexvirus / Q000235:genetics / Q000302:isolation & purification*; D014709:Vero Cells; D014773:Virology / Q000379:methods*
D004279:DNA, Viral