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

Adrian Recinos; R. Stephen Lloyd

doi:10.1016/S0006-291X(86)80587-3

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

Adrian Recinos, R. Stephen Lloyd

Research output: Contribution to journal › Article › peer-review

18 Scopus citations

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 language	English (US)
Pages (from-to)	945-952
Number of pages	8
Journal	Biochemical and Biophysical Research Communications
Volume	138
Issue number	2
DOIs	https://doi.org/10.1016/S0006-291X(86)80587-3
State	Published - Jul 31 1986
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/S0006-291X(86)80587-3

Cite this

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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.",

author = "Adrian Recinos and Lloyd, {R. Stephen}",

note = "Funding Information: To the best of our knowledge, the concept of using a bridge oligonucleotide probe was developed in 1982 within the Gram-negative Expression Group of the Genex Corporation, Gaithersberg MD. Contributing scientists were Drs. Jim Anderson, Dorthea Scandella, Stephen Lloyd, Patrick Arthur and Ethyl 3ackson. We wish to thank Ms. Doris Harris for her efficient preparation of this manuscript. This research was supported by NIH ES 00267 and NIH ES 07028. A.R. is a predoctoral fellow supported by NIH ES 07028.",

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doi = "10.1016/S0006-291X(86)80587-3",

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AU - Recinos, Adrian

AU - Lloyd, R. Stephen

N1 - Funding Information: To the best of our knowledge, the concept of using a bridge oligonucleotide probe was developed in 1982 within the Gram-negative Expression Group of the Genex Corporation, Gaithersberg MD. Contributing scientists were Drs. Jim Anderson, Dorthea Scandella, Stephen Lloyd, Patrick Arthur and Ethyl 3ackson. We wish to thank Ms. Doris Harris for her efficient preparation of this manuscript. This research was supported by NIH ES 00267 and NIH ES 07028. A.R. is a predoctoral fellow supported by NIH ES 07028.

PY - 1986/7/31

Y1 - 1986/7/31

N2 - 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.

AB - 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.

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

Abstract

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