Spectral genotyping of human alleles

Science

Volumn 279, Issue 5354, 1998, Pages 1228-1229

  Kostrikis, Leondios G ^a   Tyagi, Sanjay ^b   Mhlanga, Musa M ^b   Ho, David D ^a   Kramer, Fred Russell ^b  

^a ROCKEFELLER UNIVERSITY   (United States)

^b PUBLIC HEALTH RESEARCH INSTITUTE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CHEMOKINE RECEPTOR; DNA; FLUORESCEIN; RHODAMINE;

ALLELE; DISEASE PREDISPOSITION; FLUORESCENCE; GENOTYPE; HUMAN; HUMAN IMMUNODEFICIENCY VIRUS INFECTION; OLIGONUCLEOTIDE PROBE; POLYMERASE CHAIN REACTION; PRIORITY JOURNAL; SHORT SURVEY;

ALLELES; COLORING AGENTS; DNA MUTATIONAL ANALYSIS; FLUORESCEIN; GENOTYPE; HETEROZYGOTE; HOMOZYGOTE; HUMANS; OLIGONUCLEOTIDE PROBES; POLYMERASE CHAIN REACTION; RECEPTORS, CCR5; RHODAMINES; SPECTROMETRY, FLUORESCENCE;

HUMAN IMMUNODEFICIENCY VIRUS;

EID: 0032549128     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.279.5354.1228     Document Type: Short Survey

References (21)

1. R. Cotton, Mutat. Detection (Oxford Univ. Press, Oxford, 1997).
2. J. Keen, D. Lester, C. Ingkehearn, A. Curtis, S. Bhattacharya, Trends Genet. 7, 5 (1991).
3. M. Orita, H. Iwahana, H. Kanazawa, K. Hayashi, T. Sekiya, Proc. Natl. Acad. Sci. U.S.A. 86, 2766 (1989).
4. R. Saiki, T. Bugawan, G. Horn, K. Mullis, H. Erlich, Nature 324, 163 (1986).
5. S. Tyagi, F. Kramer, Nat. Biotechnol. 14, 303 (1996).
6. S. Tyagi, D. Bratu, F. Kramer, ibid. 16, 49 (1998).
7. R. Liu et al., Cell 86, 367 (1996).
8. M. Samson et al., Nature 382, 722 (1996).
9. M. Dean et al., Science 273, 1856 (1996).
10. Y. Huang et al., Nat. Med. 2, 1240 (1996).
11. J. Rappaport et al., Lancet 349, 922 (1997).
12. The sequence of the wild-type-specific molecular beacon was fluorescein-5′-CCGGTCTG-GAAATTCTTCCAGAATTGATACTGACCGG-3′-DABCYL and the sequence of the mutant-specific molecular beacon was tetramethylrhodamine-5′-CGGCTATCTTTAATGTATGGAAAATGAGAGCCG-3'-DABCYL, where underlines identify the complementary sequences that form the stem and DABCYL is the quencher 4-(4′-dimethylaminophenylazo)benzoic acid. A protocol for synthesizing molecular beacons is available on the World Wide Web at www.phri.nyu.edu/ molecular_beacons.
13. 2, and 10 mM tris-HCl (pH 8.3). Forty cycles of amplification (94°C denaturation for 30 s, 6O°C annealing for 60 s, and 72°C polymerization for 30 s) were performed in a spectrofluorometric thermal cycler possessing a 488-nm laser light source.
14. C. Heid, J. Stevens, K Livak, P. Williams, Genome Res. 6, 986 (1996).
15. C. Cantor, Nat. Biotechnol. 14, 264 (1996).
16. G. Bonnet, S. Tyagi, F. Kramer, A. Libchaber, in preparation.
17. L. Kostrikis et al., Nat. Med., in press.
18. M. Smith et al., Science 277, 959 (1997).
19. R. Ehricht, T. Kirner, T. Ellinger, P. Foerster, S. McCaskill, Nucleic Acids Res. 25, 4697 (1997).
20. T. Matsuo, Biochim. Biophys. Acta, in press.
21. Supported by the Aaron Diamond Foundation and NIH (grant HL-43521). We thank W. Paxton, Y. Huang, and L. Zhang for providing samples of human DNA. L. Kostrikis is an Aaron Diamond Fellow.