A quantitative polymerase chain reaction method for determining copy number within the Prader-Willi/Angelman syndrome critical region
A quantitative polymerase chain reaction method for determining copy number within the Prader-Willi/Angelman syndrome critical region
The Prader–Willi/Angelman syndrome critical region (PWACR) on 15q11-q13 is prone to structural rearrangements, including deletions (1), duplications (2) and supernumerary marker chromosomes [SMC(15)s] (3), which can give rise to a variety of phenotypic effects dependent on the origin and nature of the rearrangement (4). A number of methods, such as cytogenetic analysis, fluorescence in situ hybridization (FISH) and methylation-specific or microsatellite polymerase chain reaction (PCR), are currently available for diagnosing these abnormalities (5). To supplement these approaches we have designed a quick and effective method for determining the copy number of the 15q11-q13 region, based upon quantitative PCR coupled to automated gene scanning. Two target sequences are amplified relative to an internal reference (6, 7) and the PCR products are then analyzed directly on an ABI 377 automated sequencer. Comparison of the ratios of reference and target sequence peaks allows the copy number of the 15q11-q13 region to be calculated.
76-78
Roberts, S.E.
1ba6b93b-6904-4bf4-b83d-600372bf6c60
Thomas, N.S.
df2d7c6d-2c96-4aaa-a7ef-7f7987759cf4
2003
Roberts, S.E.
1ba6b93b-6904-4bf4-b83d-600372bf6c60
Thomas, N.S.
df2d7c6d-2c96-4aaa-a7ef-7f7987759cf4
Roberts, S.E. and Thomas, N.S.
(2003)
A quantitative polymerase chain reaction method for determining copy number within the Prader-Willi/Angelman syndrome critical region.
Clinical Genetics, 64 (1), .
(doi:10.1034/j.1399-0004.2003.00094.x).
Abstract
The Prader–Willi/Angelman syndrome critical region (PWACR) on 15q11-q13 is prone to structural rearrangements, including deletions (1), duplications (2) and supernumerary marker chromosomes [SMC(15)s] (3), which can give rise to a variety of phenotypic effects dependent on the origin and nature of the rearrangement (4). A number of methods, such as cytogenetic analysis, fluorescence in situ hybridization (FISH) and methylation-specific or microsatellite polymerase chain reaction (PCR), are currently available for diagnosing these abnormalities (5). To supplement these approaches we have designed a quick and effective method for determining the copy number of the 15q11-q13 region, based upon quantitative PCR coupled to automated gene scanning. Two target sequences are amplified relative to an internal reference (6, 7) and the PCR products are then analyzed directly on an ABI 377 automated sequencer. Comparison of the ratios of reference and target sequence peaks allows the copy number of the 15q11-q13 region to be calculated.
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Published date: 2003
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Local EPrints ID: 24922
URI: http://eprints.soton.ac.uk/id/eprint/24922
ISSN: 0009-9163
PURE UUID: 0b75506a-8ca9-4a99-90df-17da545a2a26
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Date deposited: 06 Apr 2006
Last modified: 15 Mar 2024 06:59
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Author:
S.E. Roberts
Author:
N.S. Thomas
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