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Genome Walking by Next Generation Sequencing Approaches

Genome Walking (GW) comprises a number of PCR-based methods for the identification of nucleotide sequences flanking known regions. The different methods have been used for several purposes: from de novo sequencing, useful for the identification of unknown regions, to the characterization of insertio...

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Detalles Bibliográficos
Autores principales: Volpicella, Mariateresa, Leoni, Claudia, Costanza, Alessandra, Fanizza, Immacolata, Placido, Antonio, Ceci, Luigi R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4009806/
https://www.ncbi.nlm.nih.gov/pubmed/24832505
http://dx.doi.org/10.3390/biology1030495
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author Volpicella, Mariateresa
Leoni, Claudia
Costanza, Alessandra
Fanizza, Immacolata
Placido, Antonio
Ceci, Luigi R.
author_facet Volpicella, Mariateresa
Leoni, Claudia
Costanza, Alessandra
Fanizza, Immacolata
Placido, Antonio
Ceci, Luigi R.
author_sort Volpicella, Mariateresa
collection PubMed
description Genome Walking (GW) comprises a number of PCR-based methods for the identification of nucleotide sequences flanking known regions. The different methods have been used for several purposes: from de novo sequencing, useful for the identification of unknown regions, to the characterization of insertion sites for viruses and transposons. In the latter cases Genome Walking methods have been recently boosted by coupling to Next Generation Sequencing technologies. This review will focus on the development of several protocols for the application of Next Generation Sequencing (NGS) technologies to GW, which have been developed in the course of analysis of insertional libraries. These analyses find broad application in protocols for functional genomics and gene therapy. Thanks to the application of NGS technologies, the original vision of GW as a procedure for walking along an unknown genome is now changing into the possibility of observing the parallel marching of hundreds of thousands of primers across the borders of inserted DNA molecules in host genomes.
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spelling pubmed-40098062014-05-07 Genome Walking by Next Generation Sequencing Approaches Volpicella, Mariateresa Leoni, Claudia Costanza, Alessandra Fanizza, Immacolata Placido, Antonio Ceci, Luigi R. Biology (Basel) Review Genome Walking (GW) comprises a number of PCR-based methods for the identification of nucleotide sequences flanking known regions. The different methods have been used for several purposes: from de novo sequencing, useful for the identification of unknown regions, to the characterization of insertion sites for viruses and transposons. In the latter cases Genome Walking methods have been recently boosted by coupling to Next Generation Sequencing technologies. This review will focus on the development of several protocols for the application of Next Generation Sequencing (NGS) technologies to GW, which have been developed in the course of analysis of insertional libraries. These analyses find broad application in protocols for functional genomics and gene therapy. Thanks to the application of NGS technologies, the original vision of GW as a procedure for walking along an unknown genome is now changing into the possibility of observing the parallel marching of hundreds of thousands of primers across the borders of inserted DNA molecules in host genomes. MDPI 2012-10-01 /pmc/articles/PMC4009806/ /pubmed/24832505 http://dx.doi.org/10.3390/biology1030495 Text en © 2012 by the authors; licensee MDPI, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0/ This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Review
Volpicella, Mariateresa
Leoni, Claudia
Costanza, Alessandra
Fanizza, Immacolata
Placido, Antonio
Ceci, Luigi R.
Genome Walking by Next Generation Sequencing Approaches
title Genome Walking by Next Generation Sequencing Approaches
title_full Genome Walking by Next Generation Sequencing Approaches
title_fullStr Genome Walking by Next Generation Sequencing Approaches
title_full_unstemmed Genome Walking by Next Generation Sequencing Approaches
title_short Genome Walking by Next Generation Sequencing Approaches
title_sort genome walking by next generation sequencing approaches
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4009806/
https://www.ncbi.nlm.nih.gov/pubmed/24832505
http://dx.doi.org/10.3390/biology1030495
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