利用结构独特的分子标识符改进了数字测序

IF 10.1 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Genome Biology Pub Date : 2025-02-25 DOI:10.1186/s13059-025-03504-x

Peter Micallef, Manuel Luna Santamaría, Mandy Escobar, Daniel Andersson, Tobias Österlund, Pia Mouhanna, Stefan Filges, Gustav Johansson, Henrik Fagman, Christoffer Vannas, Anders Ståhlberg

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引用次数: 0

摘要

数字测序使用独特的分子标识符（UMIs）来纠正聚合酶诱导的错误和扩增偏差。在这里，我们设计了19种不同结构的umi，以最大限度地减少在使用SiMSen-Seq构建文库期间引物形成非特异性PCR产物的能力，SiMSen-Seq是一种基于PCR的数字测序方法，具有灵活的多路复用能力，适用于肿瘤突变分析。与非结构化参考UMI相比，所有结构化UMI设计都显示出更高的分析性能。性能最好的结构化UMI设计在分析和测序性能的所有测试方面都有显着改善，能够可靠地检测低变异等位基因频率。

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Digital sequencing is improved by using structured unique molecular identifiers

Digital sequencing uses unique molecular identifiers (UMIs) to correct for polymerase induced errors and amplification biases. Here, we design 19 different structured UMIs to minimize the capacity of primers to form non-specific PCR products during library construction using SiMSen-Seq, a PCR-based digital sequencing approach with flexible multiplexing capabilities suitable for tumor-informed mutation analysis. All structured UMI designs demonstrate enhanced assay performance compared with an unstructured reference UMI. The best performing structured UMI design shows significant improvements in all tested aspects of assay and sequencing performance with the ability to reliable detect low variant allele frequencies.

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来源期刊

Genome Biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

21.00

自引率

3.30%

发文量

241

审稿时长

2 months

期刊介绍： Genome Biology stands as a premier platform for exceptional research across all domains of biology and biomedicine, explored through a genomic and post-genomic lens. With an impressive impact factor of 12.3 (2022),* the journal secures its position as the 3rd-ranked research journal in the Genetics and Heredity category and the 2nd-ranked research journal in the Biotechnology and Applied Microbiology category by Thomson Reuters. Notably, Genome Biology holds the distinction of being the highest-ranked open-access journal in this category. Our dedicated team of highly trained in-house Editors collaborates closely with our esteemed Editorial Board of international experts, ensuring the journal remains on the forefront of scientific advances and community standards. Regular engagement with researchers at conferences and institute visits underscores our commitment to staying abreast of the latest developments in the field.