Best Practices for Illumina Library Preparation.
Best Practices for Illumina Library Preparation.
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DOI：
10.1002/cphg.86
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发表时间：
2019-06-01
影响因子：
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通讯作者：
Quail, Michael A
中科院分区：
文献类型：
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作者：
Bronner, Iraad F;Quail, Michael A
在本单元中，我们描述了一组用于Illumina文库制备的方案和建议。我们回顾了模板定量方法的最佳实践;模板片段化方法;固相反向固定化清除，包括缓冲液交换和大小选择;末端修复，A-加尾和接头连接;索引策略;关于是否使用聚合酶链反应的考虑;最终文库定量方法;以及标准化和合并策略。这些工作流程适用于高通量和低通量Illumina文库制备，应有助于减少偏倚，提高成本效益，并产生高文库产量。这是本单元以前版本的广泛更新。John Wiley & Sons，Inc._关于我们
In this unit, we describe a set of protocols and recommendations for Illumina library preparation. We review best practices in template quantitation methods; template fragmentation methodologies; solid-phase reverse-immobilization cleanup, including buffer exchange and size selection; end repair, A-tailing, and adapter ligation; indexing strategies; considerations regarding whether to use polymerase chain reaction; final library quantification methodologies; and normalization and pooling strategies. These workflows are applicable to both high-throughput and low-throughput Illumina library preparation and should help reduce bias, increase cost effectiveness, and produce high library yields. This is an extensive update of the previous version of this unit. © 2019 by John Wiley & Sons, Inc.