Extraction of Cell-Associated Varicella-Zoster Virus DNA With Triton X-100-NaCl

J Virol Methods. 1982 May;4(4-5):263-75. doi: 10.1016/0166-0934(82)90073-8.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Centrifugation, Isopycnic
  • Chlorocebus aethiops
  • DNA Restriction Enzymes
  • DNA, Viral / isolation & purification*
  • Herpesvirus 3, Human / analysis*
  • Microscopy, Electron
  • Molecular Weight
  • Octoxynol
  • Polyethylene Glycols
  • Sodium Chloride

Substances

  • DNA, Viral
  • Polyethylene Glycols
  • Sodium Chloride
  • Octoxynol
  • DNA Restriction Enzymes