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g you The direct determination of haplotypes from extended regions of genomic DNA
BACKGROUND: One of the major obstacles to the exploitation of genetic variation in human medicine, veterinary medicine, and animal breeding is the difficulty in defining haplotypes in unrelated individuals. RESULTS: We have developed a Multiplex Double Amplification Refractory Mutation System combin...
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2996965/ https://www.ncbi.nlm.nih.gov/pubmed/20370899 http://dx.doi.org/10.1186/1471-2164-11-223 |
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author | Stirling, David Stear, Michael J |
author_facet | Stirling, David Stear, Michael J |
author_sort | Stirling, David |
collection | PubMed |
description | BACKGROUND: One of the major obstacles to the exploitation of genetic variation in human medicine, veterinary medicine, and animal breeding is the difficulty in defining haplotypes in unrelated individuals. RESULTS: We have developed a Multiplex Double Amplification Refractory Mutation System combined with Solid Phase PCR on Fluorescently labelled beads. The process is inherently amenable to automation. It provides a high degree of internal Quality Control, as each PCR product is represented in duplicate on the bead array, and each SNP is tested against multiple partners. This technique can resolve very complex genotypes into their constituent haplotypes; it defined all the alleles at 60 SNP in exon 2 of the ovine DRB1 MHC locus in a sample of 109 rams. These 60 SNP formed 33 DRB1 exon 2 alleles; two of which had not been previously identified; although both of them have been independently confirmed. CONCLUSION: This technique has the same resolution as allele specific sequencing. Sequencing has the advantage of identifying novel polymorphic sites but where all SNP sites have been identified this novel procedure can resolve all alleles and haplotypes and identify novel combinations of polymorphisms. This method is similar in price to direct sequencing and provides a low cost system for direct haplotyping of extended DNA sequences. |
format | Text |
id | pubmed-2996965 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-29969652010-12-07 g you The direct determination of haplotypes from extended regions of genomic DNA Stirling, David Stear, Michael J BMC Genomics Methodology Article BACKGROUND: One of the major obstacles to the exploitation of genetic variation in human medicine, veterinary medicine, and animal breeding is the difficulty in defining haplotypes in unrelated individuals. RESULTS: We have developed a Multiplex Double Amplification Refractory Mutation System combined with Solid Phase PCR on Fluorescently labelled beads. The process is inherently amenable to automation. It provides a high degree of internal Quality Control, as each PCR product is represented in duplicate on the bead array, and each SNP is tested against multiple partners. This technique can resolve very complex genotypes into their constituent haplotypes; it defined all the alleles at 60 SNP in exon 2 of the ovine DRB1 MHC locus in a sample of 109 rams. These 60 SNP formed 33 DRB1 exon 2 alleles; two of which had not been previously identified; although both of them have been independently confirmed. CONCLUSION: This technique has the same resolution as allele specific sequencing. Sequencing has the advantage of identifying novel polymorphic sites but where all SNP sites have been identified this novel procedure can resolve all alleles and haplotypes and identify novel combinations of polymorphisms. This method is similar in price to direct sequencing and provides a low cost system for direct haplotyping of extended DNA sequences. BioMed Central 2010-04-06 /pmc/articles/PMC2996965/ /pubmed/20370899 http://dx.doi.org/10.1186/1471-2164-11-223 Text en Copyright ©2010 Stirling and Stear; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Article Stirling, David Stear, Michael J g you The direct determination of haplotypes from extended regions of genomic DNA |
title | g you The direct determination of haplotypes from extended regions of genomic DNA |
title_full | g you The direct determination of haplotypes from extended regions of genomic DNA |
title_fullStr | g you The direct determination of haplotypes from extended regions of genomic DNA |
title_full_unstemmed | g you The direct determination of haplotypes from extended regions of genomic DNA |
title_short | g you The direct determination of haplotypes from extended regions of genomic DNA |
title_sort | g you the direct determination of haplotypes from extended regions of genomic dna |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2996965/ https://www.ncbi.nlm.nih.gov/pubmed/20370899 http://dx.doi.org/10.1186/1471-2164-11-223 |
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