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Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array
The scarce amount of DNA contained in a singe cell is a limiting factor for clinical application of preimplantation genetic diagnosis mainly due to the risk of misdiagnosis caused by allele dropout and the difficulty in obtaining copy number variations in all 23 pairs of chromosomes. Multiple displa...
Autores principales: | , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2009
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2762374/ https://www.ncbi.nlm.nih.gov/pubmed/19671595 http://dx.doi.org/10.1093/molehr/gap066 |
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author | Ling, Jiawei Zhuang, Guanglun Tazon-Vega, Barbara Zhang, Chenhui Cao, Baoqiang Rosenwaks, Zev Xu, Kangpu |
author_facet | Ling, Jiawei Zhuang, Guanglun Tazon-Vega, Barbara Zhang, Chenhui Cao, Baoqiang Rosenwaks, Zev Xu, Kangpu |
author_sort | Ling, Jiawei |
collection | PubMed |
description | The scarce amount of DNA contained in a singe cell is a limiting factor for clinical application of preimplantation genetic diagnosis mainly due to the risk of misdiagnosis caused by allele dropout and the difficulty in obtaining copy number variations in all 23 pairs of chromosomes. Multiple displacement amplification (MDA) has been reported to generate large quantity of products from small amount of templates. Here, we evaluated the fidelity of whole-genome amplification MDA from single or a few cells and determined the accuracy of chromosome copy number assessment on these MDA products using an Affymetrix 10K 2.0 SNP Mapping Array. An average coverage rate (86.2%) from single cells was obtained and the rates increased significantly when five or more cells were used as templates. Higher concordance for chromosome copy number from single cells could be achieved when the MDA amplified product was used as reference (93.1%) than when gDNA used as reference (82.8%). The present study indicates that satisfactory genome coverage can be obtained from single-cell MDA which may be used for studies where only a minute amount of genetic materials is available. Clinically, MDA coupled with SNP mapping array may provide a reliable and accurate method for chromosome copy number analysis and most likely for the detection of single-gene disorders as well. |
format | Text |
id | pubmed-2762374 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-27623742009-10-16 Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array Ling, Jiawei Zhuang, Guanglun Tazon-Vega, Barbara Zhang, Chenhui Cao, Baoqiang Rosenwaks, Zev Xu, Kangpu Mol Hum Reprod Original Research The scarce amount of DNA contained in a singe cell is a limiting factor for clinical application of preimplantation genetic diagnosis mainly due to the risk of misdiagnosis caused by allele dropout and the difficulty in obtaining copy number variations in all 23 pairs of chromosomes. Multiple displacement amplification (MDA) has been reported to generate large quantity of products from small amount of templates. Here, we evaluated the fidelity of whole-genome amplification MDA from single or a few cells and determined the accuracy of chromosome copy number assessment on these MDA products using an Affymetrix 10K 2.0 SNP Mapping Array. An average coverage rate (86.2%) from single cells was obtained and the rates increased significantly when five or more cells were used as templates. Higher concordance for chromosome copy number from single cells could be achieved when the MDA amplified product was used as reference (93.1%) than when gDNA used as reference (82.8%). The present study indicates that satisfactory genome coverage can be obtained from single-cell MDA which may be used for studies where only a minute amount of genetic materials is available. Clinically, MDA coupled with SNP mapping array may provide a reliable and accurate method for chromosome copy number analysis and most likely for the detection of single-gene disorders as well. Oxford University Press 2009-11 2009-08-11 /pmc/articles/PMC2762374/ /pubmed/19671595 http://dx.doi.org/10.1093/molehr/gap066 Text en © The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Research Ling, Jiawei Zhuang, Guanglun Tazon-Vega, Barbara Zhang, Chenhui Cao, Baoqiang Rosenwaks, Zev Xu, Kangpu Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array |
title | Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array |
title_full | Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array |
title_fullStr | Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array |
title_full_unstemmed | Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array |
title_short | Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array |
title_sort | evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by snp array |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2762374/ https://www.ncbi.nlm.nih.gov/pubmed/19671595 http://dx.doi.org/10.1093/molehr/gap066 |
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