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Titolo:
Homogeneous assays for single-nucleotide polymorphism typing using AlphaScreen
Autore:
Beaudet, L; Bedard, J; Breton, B; Mercuri, RJ; Budarf, ML;
Indirizzi:
BioSignal Packard Inc, Montreal, PQ H3J 1R4, Canada BioSignal Packard IncMontreal PQ Canada H3J 1R4 real, PQ H3J 1R4, Canada Univ Penn, Sch Med, Dept Pediat, Div Human Genet, Philadelphia, PA 19104 USA Univ Penn Philadelphia PA USA 19104 man Genet, Philadelphia, PA 19104 USA
Titolo Testata:
GENOME RESEARCH
fascicolo: 4, volume: 11, anno: 2001,
pagine: 600 - 608
SICI:
1088-9051(200104)11:4<600:HAFSPT>2.0.ZU;2-X
Fonte:
ISI
Lingua:
ENG
Soggetto:
OXYGEN CHANNELING ASSAY; TOF MASS-SPECTROMETRY; MOLECULAR BEACONS; OLIGONUCLEOTIDE ARRAYS; ALZHEIMERS-DISEASE; PRIMER EXTENSION; HUMAN GENOME; DNA; ALLELE; PHARMACOGENOMICS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
42
Recensione:
Indirizzi per estratti:
Indirizzo: Beaudet, L BioSignal Packard Inc, Montreal, PQ H3J 1R4, Canada BioSignal Packard Inc Montreal PQ Canada H3J 1R4 J 1R4, Canada
Citazione:
L. Beaudet et al., "Homogeneous assays for single-nucleotide polymorphism typing using AlphaScreen", GENOME RES, 11(4), 2001, pp. 600-608

Abstract

AlphaScreen technology allows the development of high-throughput homogeneous proximity assays. In these assays, signal is generated when 680 nm laserfight irradiates a donor bead in close proximity to an acceptor bead. For the detection of nucleic acids, donor and acceptor beads are brought into proximity by two bridging probes that hybridize simultaneously to a common target and to the generic oligonucleotides attached covalently to the beads. This method allows the detection of as little as 10 amole of a single-stranded DNA target. The combination of AlphaScreen with allele-specific amplification (ASA) and allele-specific hybridization (ASH) has allowed the development of two homogenous single-nucleotide polymorphism (SNP) genotyping platforms. Both types of assay are very robust, routinely giving accurate genotyping results with <2 ng of genomic DNA per genotype. An AlphaScreen validation study was performed for 12 SNPs by using ASA assays and seven SNPs by using ASH assays. More than 580 samples were genotyped with accuracy >99%. The two assays are remarkably simple, requiring no post-PCR manipulations. Genotyping has been performed successfully in 96- and 384-well formats with volumes as small as 2 mul, allowing a considerable reduction in the amount of reagents and genomic DNA necessary for genotyping. These results showthat the AlphaScreen technology can be successfully adapted to high-throughput genotyping.

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Documento generato il 06/04/20 alle ore 01:38:19