Fosmid libraries for genomic structural variation detection.
Fosmid libraries for genomic structural variation detection.
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DOI：
10.1002/0471142905.hg0520s54
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发表时间：
2007-07-01
影响因子：
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通讯作者：
Ebling, Heather M
中科院分区：
文献类型：
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作者：
Donahue, William F;Ebling, Heather M
Fosmid库已经证明了它们在许多应用程序中的实用性。这些包括在序列组装中填充BAC和小插入文库之间的缺口，进行杂交/筛选研究以分离基因组内的功能元件（Vergin等人，1998），以及在结构变异研究中检测插入、缺失和重排（Tuzun等人，2005年）。本单元涵盖了构建fosmid库的基本方法，这些库具有适合这些应用的紧密插入尺寸。基本方案1涵盖了剪切、大小选择和从脉冲场凝胶中回收DNA。基本方案2包括将插入DNA克隆到fosmid载体中，将DNA包装到感染性噬菌体颗粒中，以及细菌的感染/转化。提供了一个评论部分，概述了fosmid库构建中涉及的许多关键参数，沿着一些额外的背景信息和一个讨论预期结果的部分。
Fosmid libraries have demonstrated their utility for a number of applications. These include filling gaps between BACs and small insert libraries in sequence assemblies, performing hybridization/screening studies to isolate functional elements within the genome (Vergin et al., 1998), and detecting insertions, deletions, and rearrangements in structural variation studies (Tuzun et al., 2005). This unit covers the basic methodologies for the construction of fosmid libraries with tight insert sizes suitable for these applications. Basic Protocol 1 covers the shearing, size selection, and recovery of DNA from a pulsed-field gel. Basic Protocol 2 covers the cloning of insert DNA into the fosmid vector, packaging of DNA into infective phage particles, and the infection/transformation of bacteria. A commentary section is provided, which outlines many of the critical parameters involved in fosmid library construction, along with some additional background information and a section discussing anticipated results.