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基于纠错 DNA 和 RNA 测序的稀有事件检测方法

JoVE 杂志
遗传学

This content is Free Access.

JoVE 杂志遗传学

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

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基于纠错 DNA 和 RNA 测序的稀有事件检测方法

DOI:

10.3791/57509-v

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10:36 min

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August 03, 2018

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Wing H. Wong*^1,2, R. Spencer Tong*^1,2, Andrew L. Young², Todd E. Druley²

¹Department of Pediatrics, Division of Hematology and Oncology,Washington University School of Medicine, ²Center for Genome Sciences and Systems Biology,Washington University School of Medicine

Chapters

00:04Title
00:33Incorporating i5 and i7 Adapters onto Error-corrected Sequencing Libraries
03:36Quantifying the Libraries with QX200 Digital Droplet PCR (ddPCR)
06:14Amplifying and Normalizing the Libraries for Sequencing
07:37Results: Mutation-calling with Error-corrected Sequencing (ECS)
09:14Conclusion

Summary

自动翻译

下一代测序 (0.5–2.0%) 是基因组特征的有力工具, 受平台高错误率的限制。我们描述了我们的错误纠正排序方法, 使我们能够避免的误差率和检测突变的变异等位基因分数罕见的0.0001。

Tags

Rare Event Detection Error-corrected DNA Sequencing Error-corrected RNA Sequencing Clonal Single-nucleotide Variants Small Indels Unique Molecular Identifiers Q5 Master Mix PCR Magnetic Bead Cleanup Illumina Chemistry Gene Panels

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