A modified method for whole exome resequencing from minimal amounts of starting DNA.

Next generation DNA sequencing (NGS) technologies have revolutionized the pace at which whole genome and exome sequences can be generated. However, despite these advances, many of the methods for targeted resequencing, such as the generation of high-depth exome sequences, are somewhat limited by the...

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Autores principales:	Iwanka Kozarewa, Juan Manuel Rosa-Rosa, Christopher P Wardell, Brian A Walker, Kerry Fenwick, Ioannis Assiotis, Costas Mitsopoulos, Marketa Zvelebil, Gareth J Morgan, Alan Ashworth, Christopher J Lord
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4990299967334bd2b847b2b716fc5096
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spelling	oai:doaj.org-article:4990299967334bd2b847b2b716fc50962021-11-18T07:26:05ZA modified method for whole exome resequencing from minimal amounts of starting DNA.1932-620310.1371/journal.pone.0032617https://doaj.org/article/4990299967334bd2b847b2b716fc50962012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22403682/?tool=EBIhttps://doaj.org/toc/1932-6203Next generation DNA sequencing (NGS) technologies have revolutionized the pace at which whole genome and exome sequences can be generated. However, despite these advances, many of the methods for targeted resequencing, such as the generation of high-depth exome sequences, are somewhat limited by the relatively large amounts of starting DNA that are normally required. In the case of tumour analysis this is particularly pertinent as many tumour biopsies often return submicrogram quantities of DNA, especially when tumours are microdissected prior to analysis. Here, we present a method for exome capture and resequencing using as little as 50 ng of starting DNA. The sequencing libraries generated by this minimal starting amount (MSA-Cap) method generate datasets that are comparable to standard amount (SA) whole exome libraries that use three micrograms of starting DNA. This method, which can be performed in most laboratories using commonly available reagents, has the potential to enhance large scale profiling efforts such as the resequencing of tumour exomes.Iwanka KozarewaJuan Manuel Rosa-RosaChristopher P WardellBrian A WalkerKerry FenwickIoannis AssiotisCostas MitsopoulosMarketa ZvelebilGareth J MorganAlan AshworthChristopher J LordPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 3, p e32617 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Iwanka Kozarewa Juan Manuel Rosa-Rosa Christopher P Wardell Brian A Walker Kerry Fenwick Ioannis Assiotis Costas Mitsopoulos Marketa Zvelebil Gareth J Morgan Alan Ashworth Christopher J Lord A modified method for whole exome resequencing from minimal amounts of starting DNA.
description	Next generation DNA sequencing (NGS) technologies have revolutionized the pace at which whole genome and exome sequences can be generated. However, despite these advances, many of the methods for targeted resequencing, such as the generation of high-depth exome sequences, are somewhat limited by the relatively large amounts of starting DNA that are normally required. In the case of tumour analysis this is particularly pertinent as many tumour biopsies often return submicrogram quantities of DNA, especially when tumours are microdissected prior to analysis. Here, we present a method for exome capture and resequencing using as little as 50 ng of starting DNA. The sequencing libraries generated by this minimal starting amount (MSA-Cap) method generate datasets that are comparable to standard amount (SA) whole exome libraries that use three micrograms of starting DNA. This method, which can be performed in most laboratories using commonly available reagents, has the potential to enhance large scale profiling efforts such as the resequencing of tumour exomes.
format	article
author	Iwanka Kozarewa Juan Manuel Rosa-Rosa Christopher P Wardell Brian A Walker Kerry Fenwick Ioannis Assiotis Costas Mitsopoulos Marketa Zvelebil Gareth J Morgan Alan Ashworth Christopher J Lord
author_facet	Iwanka Kozarewa Juan Manuel Rosa-Rosa Christopher P Wardell Brian A Walker Kerry Fenwick Ioannis Assiotis Costas Mitsopoulos Marketa Zvelebil Gareth J Morgan Alan Ashworth Christopher J Lord
author_sort	Iwanka Kozarewa
title	A modified method for whole exome resequencing from minimal amounts of starting DNA.
title_short	A modified method for whole exome resequencing from minimal amounts of starting DNA.
title_full	A modified method for whole exome resequencing from minimal amounts of starting DNA.
title_fullStr	A modified method for whole exome resequencing from minimal amounts of starting DNA.
title_full_unstemmed	A modified method for whole exome resequencing from minimal amounts of starting DNA.
title_sort	modified method for whole exome resequencing from minimal amounts of starting dna.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/4990299967334bd2b847b2b716fc5096
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A modified method for whole exome resequencing from minimal amounts of starting DNA.

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