Direct cloning of unmodified PCR products by exploiting an engineered restriction site

Alessandro Testori, Irving Listowsky, Paul Sollitti

Research output: Contribution to journalArticle

Abstract

A method is described for the direct cloning of DNA fragments amplified by the polymerase chain reaction (PCR). An oligodeoxyribonucleotide, bearing two engineered XcmI sites placed in tandem, was used to generate cloning vectors bearing single 3' deoxythymidine (dT) overhangs at their ends. These 3' dT overhangs are compatible with the 3' deoxyadenosine overhangs found on most Taq polymerase-amplified PCR products. Consequently, Taq polymeraseamplified PCR products can be ligated directly into these modified restriction sites.

Original languageEnglish
Pages (from-to)151-152
Number of pages2
JournalGene
Volume143
Issue number1
DOIs
Publication statusPublished - May 27 1994

Keywords

  • dT-tailing
  • Escherichia coli
  • oligodeoxyribonucleotides
  • T vectors
  • Taq polymerase

ASJC Scopus subject areas

  • Genetics

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