Silver staining of DNA restriction fragments for the ultrarapid identification of pseudorabies virus strains.
Restriction endonuclease analysis of pseudorabies virus DNA has been used to study various virus strains. To make use of a rapid technique for the identification of viral strains, studies have been undertaken to facilitate purification of the DNA from viral particles present in cytoplasmic fractions. The ultrasensitive photochemical silver staining of nucleic acid, described by Beidler et al. (Analytic Biochemistry 1982: 126) has been adapted and applied to the detection of pseudorabies virus restriction fragments. In a period of 5 h more than 1 microgram of DNA can be extracted from a 25 cm2 plastic flask containing infected cells and purified without ultra centrifugation. Low molecular weight DNA was separated from high molecular weight DNA by polyacrylamide gel electrophoresis. The restriction fragments were selectively visualized by silver staining which can detect 0.025 micrograms of total pseudorabies virus DNA. The electrophoretic DNA pattern of vaccine and wild strains has been studied using these techniques and the results agree with previously published data.
Jestin
A
A
Ministère de l'Agriculture, Services Vétérinaires, Station de Pathologie Porcine, Ploufragan, France.
Blanchard
P
P
Vannier
P
P
Garbar-Chenon
A
A
Nicolas
J C
JC
D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004996:Ethidium; D011558:Herpesvirus 1, Suid / Q000302:isolation & purification*; D012834:Silver; D013194:Staining and Labeling / Q000379:methods*
D004279:DNA, Viral; D012834:Silver; D004262:DNA Restriction Enzymes; D004996:Ethidium