Center for Genomic Science, Istituto Italiano di Tecnologia, Milano, Italy.
European Molecular Biology Laboratory, European Bioinformatics Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK.
Methods Mol Biol. 2021;2284:569-578. doi: 10.1007/978-1-0716-1307-8_31.
The recent advent of Nanopore sequencing allows for the sequencing of full-length RNA or cDNA molecules. This new type of data introduces new challenges from the computational point of view, and requires new software as well as dedicated analysis pipelines. In this chapter, we guide the reader through the typical analysis steps required to process the raw data produced by the instrument into a table of counts suitable for downstream analyses. We first describe the procedure to convert raw direct RNA-Seq and cDNA-Seq data into sequences in fastq format. We then outline how to perform quality control and filtering steps and how to map the filtered long reads to a reference transcriptome or genome.
最近出现的纳米孔测序技术允许对全长 RNA 或 cDNA 分子进行测序。从计算的角度来看,这种新型数据带来了新的挑战,需要新的软件以及专门的分析管道。在本章中,我们将引导读者完成将仪器生成的原始数据处理成适合下游分析的计数表所需的典型分析步骤。我们首先描述将原始直接 RNA-Seq 和 cDNA-Seq 数据转换为 fastq 格式序列的过程。然后,我们概述了如何执行质量控制和过滤步骤,以及如何将过滤后的长读段映射到参考转录组或基因组上。