Abstract
Bulk RNA-sequencing (RNA-seq) is commonly used for identifying and characterizing genes through annotation, phylogeny, and differential expression analysis. For organisms without a reference genome, the need to generate a de novo transcriptome assembly for analyzing these data can prove challenging, as it is a multi-step process requiring many tools and computational resources. Therefore, a standardized and reproducible workflow using current best practices is required. We introduce the nf-core/denovotranscript workflow, an open-source solution built using Nextflow and the nf-core framework. The protocols here describe how the workflow can be used to perform pre-processing, de novo transcriptome assembly, redundancy reduction, assembly quality assessment, and quantification using RNA-seq data. This workflow offers simple installation and thorough documentation. The use of Docker, Singularity, and Podman container technologies also makes it portable across various computing environments.
| Original language | English |
|---|---|
| Article number | e70248 |
| Journal | Current Protocols |
| Volume | 5 |
| Issue number | 11 |
| DOIs | |
| State | Published - 1 Nov 2025 |
| Externally published | Yes |
Keywords
- Nextflow
- assembly
- non-model
- pipeline
- transcriptome
ASJC Scopus subject areas
- General Neuroscience
- General Immunology and Microbiology
- General Biochemistry, Genetics and Molecular Biology
- General Pharmacology, Toxicology and Pharmaceutics
- Health Informatics
- Medical Laboratory Technology
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