Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition

We characterize and extend a highly efficient method for constructing shotgun fragment libraries in which transposase catalyzes in vitro DNA fragmentation and adaptor incorporation simultaneously. We apply this method to sequencing a human genome and find that coverage biases are comparable to those...

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Detalles Bibliográficos
Autores principales: Adey, Andrew, Morrison, Hilary G, Asan, Xun, Xu, Kitzman, Jacob O, Turner, Emily H, Stackhouse, Bethany, MacKenzie, Alexandra P, Caruccio, Nicholas C, Zhang, Xiuqing, Shendure, Jay
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Method
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3046479/
https://www.ncbi.nlm.nih.gov/pubmed/21143862
http://dx.doi.org/10.1186/gb-2010-11-12-r119
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author Adey, Andrew
Morrison, Hilary G
Asan
Xun, Xu
Kitzman, Jacob O
Turner, Emily H
Stackhouse, Bethany
MacKenzie, Alexandra P
Caruccio, Nicholas C
Zhang, Xiuqing
Shendure, Jay
author_facet Adey, Andrew
Morrison, Hilary G
Asan
Xun, Xu
Kitzman, Jacob O
Turner, Emily H
Stackhouse, Bethany
MacKenzie, Alexandra P
Caruccio, Nicholas C
Zhang, Xiuqing
Shendure, Jay
author_sort Adey, Andrew
collection PubMed
description We characterize and extend a highly efficient method for constructing shotgun fragment libraries in which transposase catalyzes in vitro DNA fragmentation and adaptor incorporation simultaneously. We apply this method to sequencing a human genome and find that coverage biases are comparable to those of conventional protocols. We also extend its capabilities by developing protocols for sub-nanogram library construction, exome capture from 50 ng of input DNA, PCR-free and colony PCR library construction, and 96-plex sample indexing.
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spelling pubmed-30464792011-03-01 Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition Adey, Andrew Morrison, Hilary G Asan Xun, Xu Kitzman, Jacob O Turner, Emily H Stackhouse, Bethany MacKenzie, Alexandra P Caruccio, Nicholas C Zhang, Xiuqing Shendure, Jay Genome Biol Method We characterize and extend a highly efficient method for constructing shotgun fragment libraries in which transposase catalyzes in vitro DNA fragmentation and adaptor incorporation simultaneously. We apply this method to sequencing a human genome and find that coverage biases are comparable to those of conventional protocols. We also extend its capabilities by developing protocols for sub-nanogram library construction, exome capture from 50 ng of input DNA, PCR-free and colony PCR library construction, and 96-plex sample indexing. BioMed Central 2010 2010-12-08 /pmc/articles/PMC3046479/ /pubmed/21143862 http://dx.doi.org/10.1186/gb-2010-11-12-r119 Text en Copyright ©2010 Adey et al.; 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 Method
Adey, Andrew
Morrison, Hilary G
Asan
Xun, Xu
Kitzman, Jacob O
Turner, Emily H
Stackhouse, Bethany
MacKenzie, Alexandra P
Caruccio, Nicholas C
Zhang, Xiuqing
Shendure, Jay
Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
title Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
title_full Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
title_fullStr Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
title_full_unstemmed Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
title_short Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
title_sort rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3046479/
https://www.ncbi.nlm.nih.gov/pubmed/21143862
http://dx.doi.org/10.1186/gb-2010-11-12-r119
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