An improved rapid method for purification of herpes simplex virus DNA using cesium trifluoroacetate.
A method for purification of herpes simplex virus DNA from cell culture is described which yields highly purified viral DNA within 8 h. The method involves the freezing and thawing of virus-infected cells followed by isopycnic centrifugation of the lysate supernatant in cesium trifluoroacetate. It was found that this method recovered DNA from most of the cell-associated virus particles in such sufficient purity that the DNA was digestible with restriction enzymes and could be used to transfect cells without the need for additional purification steps. Purification of viral DNA from cells that were not subjected to freezing and thawing was less efficient due to the amount of viral DNA that remained cell-associated.
Ling
J Y
JY
Division of Molecular Virology, Baylor College of Medicine, Houston, TX, USA.
Kienzle
T E
TE
Stroop
W G
WG
D000818:Animals; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D006224:Cricetinae; D004279:DNA, Viral / Q000302:isolation & purification*; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006801:Humans; D013997:Time Factors; D014269:Trifluoroacetic Acid / Q000737:chemistry*
D004279:DNA, Viral; D014269:Trifluoroacetic Acid