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Whole genome amplification of degraded and nondegraded DNA for forensic purposes
Degraded DNA is often analyzed in forensic genetics laboratories. Reliable analysis of degraded DNA is of great importance, since its results impact the quality and reliability of expert testimonies. Recently, a number of whole genome amplification (WGA) methods have been proposed as preamplificatio...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer-Verlag
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3578730/ https://www.ncbi.nlm.nih.gov/pubmed/22940764 http://dx.doi.org/10.1007/s00414-012-0764-9 |
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author | Maciejewska, Agnieszka Jakubowska, Joanna Pawłowski, Ryszard |
author_facet | Maciejewska, Agnieszka Jakubowska, Joanna Pawłowski, Ryszard |
author_sort | Maciejewska, Agnieszka |
collection | PubMed |
description | Degraded DNA is often analyzed in forensic genetics laboratories. Reliable analysis of degraded DNA is of great importance, since its results impact the quality and reliability of expert testimonies. Recently, a number of whole genome amplification (WGA) methods have been proposed as preamplification tools. They work on the premise of being able to generate microgram quantities of DNA from as little as the quantity of DNA from a single cell. We chose, investigated, and compared seven WGA methods to evaluate their ability to “recover” degraded and nondegraded DNA: degenerate oligonucleotide-primed PCR, primer extension preamplification PCR, GenomePlex™ WGA commercial kit (Sigma), multiple displacement amplification, GenomiPhi™ Amplification kit (Amersham Biosciences), restriction and circularization-aided rolling circle amplification, and blunt-end ligation-mediated WGA. The efficiency and reliability of those methods were analyzed and compared using SGMPlus, YFiler, mtDNA, and Y-chromosome SNP typing. The best results for nondegraded DNA were obtained with GenomiPhi and PEP methods. In the case of degraded DNA (200 bp), the best results were obtained with GenomePlex which successfully amplified also severely degraded DNA (100 bp), thus enabling correct typing of mtDNA and Y-SNP loci. WGA may be very useful in analysis of low copy number DNA or degraded DNA in forensic genetics, especially after introduction of some improvements (sample pooling and replicate DNA typing). ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00414-012-0764-9) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-3578730 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Springer-Verlag |
record_format | MEDLINE/PubMed |
spelling | pubmed-35787302013-02-26 Whole genome amplification of degraded and nondegraded DNA for forensic purposes Maciejewska, Agnieszka Jakubowska, Joanna Pawłowski, Ryszard Int J Legal Med Original Article Degraded DNA is often analyzed in forensic genetics laboratories. Reliable analysis of degraded DNA is of great importance, since its results impact the quality and reliability of expert testimonies. Recently, a number of whole genome amplification (WGA) methods have been proposed as preamplification tools. They work on the premise of being able to generate microgram quantities of DNA from as little as the quantity of DNA from a single cell. We chose, investigated, and compared seven WGA methods to evaluate their ability to “recover” degraded and nondegraded DNA: degenerate oligonucleotide-primed PCR, primer extension preamplification PCR, GenomePlex™ WGA commercial kit (Sigma), multiple displacement amplification, GenomiPhi™ Amplification kit (Amersham Biosciences), restriction and circularization-aided rolling circle amplification, and blunt-end ligation-mediated WGA. The efficiency and reliability of those methods were analyzed and compared using SGMPlus, YFiler, mtDNA, and Y-chromosome SNP typing. The best results for nondegraded DNA were obtained with GenomiPhi and PEP methods. In the case of degraded DNA (200 bp), the best results were obtained with GenomePlex which successfully amplified also severely degraded DNA (100 bp), thus enabling correct typing of mtDNA and Y-SNP loci. WGA may be very useful in analysis of low copy number DNA or degraded DNA in forensic genetics, especially after introduction of some improvements (sample pooling and replicate DNA typing). ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00414-012-0764-9) contains supplementary material, which is available to authorized users. Springer-Verlag 2012-09-01 2013 /pmc/articles/PMC3578730/ /pubmed/22940764 http://dx.doi.org/10.1007/s00414-012-0764-9 Text en © The Author(s) 2012 https://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. |
spellingShingle | Original Article Maciejewska, Agnieszka Jakubowska, Joanna Pawłowski, Ryszard Whole genome amplification of degraded and nondegraded DNA for forensic purposes |
title | Whole genome amplification of degraded and nondegraded DNA for forensic purposes |
title_full | Whole genome amplification of degraded and nondegraded DNA for forensic purposes |
title_fullStr | Whole genome amplification of degraded and nondegraded DNA for forensic purposes |
title_full_unstemmed | Whole genome amplification of degraded and nondegraded DNA for forensic purposes |
title_short | Whole genome amplification of degraded and nondegraded DNA for forensic purposes |
title_sort | whole genome amplification of degraded and nondegraded dna for forensic purposes |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3578730/ https://www.ncbi.nlm.nih.gov/pubmed/22940764 http://dx.doi.org/10.1007/s00414-012-0764-9 |
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