Competitive hybridization technique

Details Competitive hybridization technique Competitive hybridization technique

1987-07-08

1996-12-10

Zitomer, Stephanie W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

935 77, 935 78, C12Q 168

Patent

active

055829708

ABSTRACT:
Discrimination between RNAs which differ by as little as a single nucleotide is accomplished by having an excess of unlabeled non-complementary oligonucleotide present during hybridization with a labeled complementary nucleotide. The non-complementary oligonucleotide blocks hybridization of the labeled complementary oligonucleotide sequence without affecting hybridization of the labeled complementary oligonucleotide to the complementary sequence. The label may be an isotope, a fluorescent group or of any other type. The technique permits examination of the transcription of highly related allelic and non-allelic genes present in the same cell and the quantification of such transcripts by use of appropriate internal control. The technique is also useful in oligonucleotide hybridization to DNA sequences which differ only by a single nucleotide.

REFERENCES:
patent: 4683194 (1987-07-01), Saiki et al.
Lathe, R., "Synthetic Oligonucleotide Probes Deduced From Amino Acid Sequence Data--Theoretical and Practical Considerations," J. Mol. Biol. 183:1-12 (1985).
Ooostra, B. A. et al., Anal. Biochem. 74:496-502 (1976).
Orkin et al., J. Clin. Invest. 71:775-779 (1983).
Wallace et al., Nuc. Acids Res. 9:879-894 (1981).
Maniatis et al., Molec Cloning:A Lab. Man. Cold Spring Harb., 1982.
Conner et al., Proc. Natl. Acad. Sci., USA, 80:278-282 (1983).
Studencki et al., Am. J. Hum. Gen. 37:42-51 (1985).
Alonso et al., Exp. Cell Res. 85:383-390 (1974).
Nozari et al., Gene 43:23-28 (1986).

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