Library preparation and data analysis packages for rapid genome sequencing.
Library preparation and data analysis packages for rapid genome sequencing.
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DOI：
10.1007/978-1-62703-122-6_1
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发表时间：
2012
期刊：
影响因子：
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通讯作者：
Freitag, Michael
中科院分区：
文献类型：
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作者：
Pomraning, Kyle R;Smith, Kristina M;Bredeweg, Erin L;Connolly, Lanelle R;Phatale, Pallavi A;Freitag, Michael
高通量测序（HTS）已迅速成为比较遗传学和基因组学的宝贵工具，现在在没有连接到大型测序中心的实验室中定期进行。在这里，我们描述了我们的协议的更新版本，用于构建单端和双端Illumina测序文库，从纯化的基因组DNA开始。本方案还可用于“多路复用”，即通过以独立于Illumina支持的方法的方式生成“条形码化”或“索引化”Illumina测序文库来分析单个流动池泳道中的若干样品。为了分析测序结果，我们提出了几种独立的方法，但最终用户应该意识到这是一个快速发展的领域，目前正在开发和测试许多比对（或“映射”）和计数算法。
High-throughput sequencing (HTS) has quickly become a valuable tool for comparative genetics and genomics and is now regularly carried out in laboratories that are not connected to large sequencing centers. Here we describe an updated version of our protocol for constructing single- and paired-end Illumina sequencing libraries, beginning with purified genomic DNA. The present protocol can also be used for “multiplexing,” i.e. the analysis of several samples in a single flowcell lane by generating “barcoded” or “indexed” Illumina sequencing libraries in a way that is independent from Illumina-supported methods. To analyze sequencing results, we suggest several independent approaches but end users should be aware that this is a quickly evolving field and that currently many alignment (or “mapping”) and counting algorithms are being developed and tested.