Genome walking by next generation sequencing approaches.
Genome walking by next generation sequencing approaches.
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DOI：
10.3390/biology1030495
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发表时间：
2012-10-01
期刊：
影响因子：
4.2
通讯作者：
Ceci LR
中科院分区：
文献类型：
--
作者：
Volpicella M;Leoni C;Costanza A;Fanizza I;Placido A;Ceci LR
基因组行走（GW）包括许多基于pcr的方法，用于鉴定已知区域两侧的核苷酸序列。不同的方法已被用于多种目的：从用于鉴定未知区域的从头测序，到病毒和转座子插入位点的表征。在后一种情况下，基因组行走方法最近通过与下一代测序技术的耦合而得到了提升。本文将重点介绍在插入文库分析过程中开发的下一代测序（NGS）技术应用于GW的几种方案。这些分析在功能基因组学和基因治疗方案中有广泛的应用。由于NGS技术的应用，最初的设想是沿着未知基因组行走的过程，现在变成了观察成千上万的引物平行穿过宿主基因组插入DNA分子边界的可能性。
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.