Rapid preparation of vaccinia virus DNA template for analysis and cloning by PCR.

Abstract

This chapter describes the preparation of template DNA from poxvirus-infected cells, plaques, or crude virus stocks for polymerase chain reaction (PCR) amplification. The advantages of this technique are that it is rapid, inexpensive, and, most importantly, reliable, requiring only centrifugation, detergent, and protease treatment. The template preparation is suitable for PCR amplification for screening viruses, cloning, transfection, and DNA sequencing.