Extraction of cell-associated varicella-zoster virus DNA with triton X-100-NaCl.
Varicella-zoster virus (VZV) DNA was extracted from infected cells with 0.25% Triton X-100-0.2 M NaCl and purified by isopycnic centrifugation in CsCl. In each of eight experiments, 1.8-9.8 micrograms VZV DNA was obtained from 107 infected cells. The VZV DNA obtained by this procedure had a molecular weight of 88-100 x 106 as determined by sucrose gradient sedimentation and electron microscopy, and cleavage patterns after digestion with four restriction enzymes that corresponded to patterns previously described with six strains of VZV; the pattern of BamHI-cleaved Triton-NaCl-extracted VZV DNA was identical to the pattern seen after DNA extraction from virions. These studies expand the usefulness of Triton X-100-NaCl for extraction of large molecular weight viral DNA from a system where considerable cell-free virus is produced (Pignatti et al., 1979, Virology 93, 260) to a system known for its marked cell association.
D000818:Animals; D002460:Cell Line; D002500:Centrifugation, Isopycnic; D002522:Chlorocebus aethiops; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000302:isolation & purification*; D014645:Herpesvirus 3, Human / Q000032:analysis*; D008854:Microscopy, Electron; D008970:Molecular Weight; D017830:Octoxynol; D011092:Polyethylene Glycols; D012965:Sodium Chloride
D004279:DNA, Viral; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D017830:Octoxynol; D004262:DNA Restriction Enzymes