Efficient screening of recombinant DNA junctions afforded by probing with synthetic "Bridge" oligonucleotides

Adrian Recinos, Robert (Stephen) Lloyd

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Procedures have been developed which simplify and expedite the screening of recombinant DNA constructions for those which only exhibit the desired DNA-DNA junctions. A synthetic DNA oligonucleotide designed to span (or "bridge") sequences around correct restriction enzyme junctions was used as a hybridization probe for the rapid identification of those sequences in several molecular cloning methodologies. It facilitated analyses of the products of random ligation reactions, as well as constructions harbored in bacteria and bacteriophage. "Bridge" probes, [32P]-end-labeled to very high specific activity, remained useful after several hybridizations and often circumvented lengthy restriction analyses.

Original languageEnglish (US)
Pages (from-to)945-952
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume138
Issue number2
DOIs
StatePublished - Jul 31 1986
Externally publishedYes

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Recombinant DNA
Oligonucleotides
Screening
DNA
Bacteriophages
Cloning
Molecular Cloning
Ligation
Bacteria
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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