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Performance of a TthPrimPol-based whole genome amplification kit for copy number alteration detection using massively parallel sequencing
Starting from only a few cells, current whole genome amplification (WGA) methods provide enough DNA to perform massively parallel sequencing (MPS). Unfortunately, all current WGA methods introduce representation bias which limits detection of copy number aberrations (CNAs) smaller than 3 Mb. A recen...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4992833/ https://www.ncbi.nlm.nih.gov/pubmed/27546482 http://dx.doi.org/10.1038/srep31825 |
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author | Deleye, Lieselot De Coninck, Dieter Dheedene, Annelies De Sutter, Petra Menten, Björn Deforce, Dieter Van Nieuwerburgh, Filip |
author_facet | Deleye, Lieselot De Coninck, Dieter Dheedene, Annelies De Sutter, Petra Menten, Björn Deforce, Dieter Van Nieuwerburgh, Filip |
author_sort | Deleye, Lieselot |
collection | PubMed |
description | Starting from only a few cells, current whole genome amplification (WGA) methods provide enough DNA to perform massively parallel sequencing (MPS). Unfortunately, all current WGA methods introduce representation bias which limits detection of copy number aberrations (CNAs) smaller than 3 Mb. A recent WGA method, called TruePrime single cell WGA, uses a recently discovered DNA primase, TthPrimPol, instead of artificial primers to initiate DNA amplification. This method could lead to a lower representation bias, and consequently to a better detection of CNAs. The enzyme requires no complementarity and thus should generate random primers, equally distributed across the genome. The performance of TruePrime WGA was assessed for aneuploidy screening and CNA analysis after MPS, starting from 1, 3 or 5 cells. Although the method looks promising, the single cell TruePrime WGA kit v1 is not suited for high resolution CNA detection after MPS because too much representation bias is introduced. |
format | Online Article Text |
id | pubmed-4992833 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-49928332016-08-30 Performance of a TthPrimPol-based whole genome amplification kit for copy number alteration detection using massively parallel sequencing Deleye, Lieselot De Coninck, Dieter Dheedene, Annelies De Sutter, Petra Menten, Björn Deforce, Dieter Van Nieuwerburgh, Filip Sci Rep Article Starting from only a few cells, current whole genome amplification (WGA) methods provide enough DNA to perform massively parallel sequencing (MPS). Unfortunately, all current WGA methods introduce representation bias which limits detection of copy number aberrations (CNAs) smaller than 3 Mb. A recent WGA method, called TruePrime single cell WGA, uses a recently discovered DNA primase, TthPrimPol, instead of artificial primers to initiate DNA amplification. This method could lead to a lower representation bias, and consequently to a better detection of CNAs. The enzyme requires no complementarity and thus should generate random primers, equally distributed across the genome. The performance of TruePrime WGA was assessed for aneuploidy screening and CNA analysis after MPS, starting from 1, 3 or 5 cells. Although the method looks promising, the single cell TruePrime WGA kit v1 is not suited for high resolution CNA detection after MPS because too much representation bias is introduced. Nature Publishing Group 2016-08-22 /pmc/articles/PMC4992833/ /pubmed/27546482 http://dx.doi.org/10.1038/srep31825 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Deleye, Lieselot De Coninck, Dieter Dheedene, Annelies De Sutter, Petra Menten, Björn Deforce, Dieter Van Nieuwerburgh, Filip Performance of a TthPrimPol-based whole genome amplification kit for copy number alteration detection using massively parallel sequencing |
title | Performance of a TthPrimPol-based whole genome amplification kit for copy number alteration detection using massively parallel sequencing |
title_full | Performance of a TthPrimPol-based whole genome amplification kit for copy number alteration detection using massively parallel sequencing |
title_fullStr | Performance of a TthPrimPol-based whole genome amplification kit for copy number alteration detection using massively parallel sequencing |
title_full_unstemmed | Performance of a TthPrimPol-based whole genome amplification kit for copy number alteration detection using massively parallel sequencing |
title_short | Performance of a TthPrimPol-based whole genome amplification kit for copy number alteration detection using massively parallel sequencing |
title_sort | performance of a tthprimpol-based whole genome amplification kit for copy number alteration detection using massively parallel sequencing |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4992833/ https://www.ncbi.nlm.nih.gov/pubmed/27546482 http://dx.doi.org/10.1038/srep31825 |
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