Characterization of Recombinant DNA Vectors by Polymerase Chain Reaction Analysis of Whole Cells

Gurpreet S. Sandhu, James W. Precup, Bruce C. Kline

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2 Scopus citations

Abstract

This chapter focuses on the characterization of recombinant DNA vector by polymerase chain reaction (PCR) analysis of whole cells. The vectors are analyzed for the presence and the orientation of the inserted sequences by restriction enzyme digestion and gel electrophoresis. Short fragments of DNA without convenient restriction sites may require DNA sequencing to obtain this information. The advantage in the use of PCR to analyze recombinant DNA molecules is that it is a rapid and reliable technique. It is a technique that can obviate the need for purification, restriction mapping, or sequencing. in terms of sensitivity, the ability to amplify DNA from single eukaryotic cells is well established. by using PCR analysis with one recombinant specific internal primer and another baculovirus-specific flanking primer, the identity of a suspected plaque can be confirmed without waiting days or weeks to be certain of visual identification.

Original languageEnglish (US)
Pages (from-to)363-368
Number of pages6
JournalMethods in Enzymology
Volume218
Issue numberC
DOIs
StatePublished - Jan 1 1993

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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