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NGSpeciesID:从长读长测序数据生成DNA条形码和扩增子共识序列。

NGSpeciesID: DNA barcode and amplicon consensus generation from long-read sequencing data.

作者信息

Sahlin Kristoffer, Lim Marisa C W, Prost Stefan

机构信息

Department of Mathematics Science for Life Laboratory Stockholm University Stockholm Sweden.

Department of Population Health and Reproduction University of California Davis CA USA.

出版信息

Ecol Evol. 2021 Jan 11;11(3):1392-1398. doi: 10.1002/ece3.7146. eCollection 2021 Feb.

Abstract

Third-generation sequencing technologies, such as Oxford Nanopore Technologies (ONT) and Pacific Biosciences (PacBio), have gained popularity over the last years. These platforms can generate millions of long-read sequences. This is not only advantageous for genome sequencing projects, but also advantageous for amplicon-based high-throughput sequencing experiments, such as DNA barcoding. However, the relatively high error rates associated with these technologies still pose challenges for generating high-quality consensus sequences. Here, we present NGSpeciesID, a program which can generate highly accurate consensus sequences from long-read amplicon sequencing technologies, including ONT and PacBio. The tool includes clustering of the reads to help filter out contaminants or reads with high error rates and employs polishing strategies specific to the appropriate sequencing platform. We show that NGSpeciesID produces consensus sequences with improved usability by minimizing preprocessing and software installation and scalability by enabling rapid processing of hundreds to thousands of samples, while maintaining similar consensus accuracy as current pipelines.

摘要

过去几年,第三代测序技术,如牛津纳米孔技术公司(ONT)和太平洋生物科学公司(PacBio)的技术,已逐渐受到欢迎。这些平台能够生成数百万条长读长序列。这不仅对基因组测序项目有利,对基于扩增子的高通量测序实验(如DNA条形码技术)也有利。然而,与这些技术相关的相对较高错误率,在生成高质量一致性序列方面仍然构成挑战。在此,我们介绍NGSpeciesID,这是一个能够从长读长扩增子测序技术(包括ONT和PacBio)生成高度准确一致性序列的程序。该工具包括对读数进行聚类,以帮助过滤掉污染物或错误率高的读数,并采用适用于相应测序平台的优化策略。我们表明,NGSpeciesID通过尽量减少预处理和软件安装,提高了一致性序列的可用性;通过能够快速处理数百至数千个样本,提高了可扩展性,同时保持与当前流程相似的一致性准确性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4d18/7863402/75f8659dc93a/ECE3-11-1392-g001.jpg

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