Oligonucleotide hybridization used to detect short non-contiguous sequences.
We describe the use of an oligonucleotide construction as a hybridization probe to detect short noncontiguous regions of sequence identity. Oligonucleotides complementary to various portions of the conserved heptamer and nonamer sequences flanking immunoglobulin variable region genes at the 3' end were used in this model system. We show that short oligonucleotides alone (7 bp or 9 bp) cannot be used as hybridization probes, but a construction containing both conserved sequences linked by a bridge will hybridize. The bridge is formed by degenerate bases (any base potentially at each position) and serves to maintain the spacing originally present between heptamer and nonamer. We show that such a bridging oligonucleotide probe can be used for hybridization analysis both on Southern blots and in bacterial screening.
Borghesi-Nicoletti
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Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore 21201.
Schulze
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D001483:Base Sequence; D015139:Blotting, Southern; D004247:DNA / Q000032:analysis; D015723:Gene Library; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D012150:Polymorphism, Restriction Fragment Length
D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D004247:DNA