Direct identification of recombinant vaccinia virus plaques by PCR.
A fast method for the identification of recombinant vaccinia viruses directly from individual plaques is described. Plaques are picked, resuspended in PBS-A and processed for PCR using two 'universal' primers. The amplified sequences are analyzed by agarose gel electrophoresis. This procedure allows discrimination between spontaneously arising TK-negative mutants, which do not carry the inserted gene, and the desired TK-negative recombinants resulting from insertional inactivation of the TK gene.
Pasamontes
L
L
F. Hoffmann-La Roche Ltd., VFRM, Basel, Switzerland.
Gubser
J
J
Wittek
R
R
Viljoen
G J
GJ
D001483:Base Sequence; D002478:Cells, Cultured; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D014616:Vaccinia virus / Q000235:genetics / Q000302:isolation & purification*; D010948:Viral Plaque Assay