Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, USA.
Center for Computational Biology, Whiting School of Engineering, Johns Hopkins University, Baltimore, MD, USA.
Nat Protoc. 2022 Dec;17(12):2815-2839. doi: 10.1038/s41596-022-00738-y. Epub 2022 Sep 28.
Metagenomic experiments expose the wide range of microscopic organisms in any microbial environment through high-throughput DNA sequencing. The computational analysis of the sequencing data is critical for the accurate and complete characterization of the microbial community. To facilitate efficient and reproducible metagenomic analysis, we introduce a step-by-step protocol for the Kraken suite, an end-to-end pipeline for the classification, quantification and visualization of metagenomic datasets. Our protocol describes the execution of the Kraken programs, via a sequence of easy-to-use scripts, in two scenarios: (1) quantification of the species in a given metagenomics sample; and (2) detection of a pathogenic agent from a clinical sample taken from a human patient. The protocol, which is executed within 1-2 h, is targeted to biologists and clinicians working in microbiome or metagenomics analysis who are familiar with the Unix command-line environment.
宏基因组实验通过高通量 DNA 测序来揭示任何微生物环境中的广泛微生物。测序数据的计算分析对于准确和完整地表征微生物群落至关重要。为了促进高效和可重复的宏基因组分析,我们介绍了 Kraken 套件的分步协议,这是一个用于分类、定量和可视化宏基因组数据集的端到端管道。我们的协议描述了通过一系列易于使用的脚本执行 Kraken 程序,有两种情况:(1) 对给定宏基因组样本中的物种进行定量;(2) 从人类患者的临床样本中检测出病原体。该协议在 1-2 小时内执行,针对的是熟悉 Unix 命令行环境的从事微生物组或宏基因组分析的生物学家和临床医生。