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Many biological and medical questions are answered based on the analysis of sequence data. However, we can find contamination, artificial spike-ins, and overrepresented rRNA (ribosomal RNA) sequences in various read collections and assemblies. In particular, spike-ins used as controls, as those known from Illumina or Nanopore data, are often not considered as contaminants and also not appropriately removed during analyses. Additionally, removing human host DNA may be necessary for data protection and ethical considerations to ensure that individuals cannot be identified. We developed CLEAN, a pipeline to remove unwanted sequences from both long- and short-read sequencing techniques. While focusing on Illumina and Nanopore data with their technology-specific control sequences, the pipeline can also be used for host decontamination of metagenomic reads and assemblies, or the removal of rRNA from RNA-Seq data. The results are the purified sequences and sequences identified as contaminated with statistics summarized in a report. The output can be used directly in subsequent analyses, resulting in faster computations and improved results. Although decontamination seems mundane, many contaminants are routinely overlooked, cleaned by steps that are not fully reproducible or difficult to trace. CLEAN facilitates reproducible, platform-independent data analysis in genomics and transcriptomics and is freely available at https://github.com/rki-mf1/clean under a BSD3 license.
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http://dx.doi.org/10.1093/nargab/lqaf105 | DOI Listing |
Microb Genom
September 2025
Department of Infection Biology, Faculty of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine, London, UK.
Amplicon sequencing is a popular method for understanding the diversity of bacterial communities in samples containing multiple organisms as exemplified by 16S rRNA sequencing. Another application of amplicon sequencing includes multiplexing both primer sets and samples, allowing sequencing of multiple targets in multiple samples in the same sequencing run. Multiple tools exist to process the amplicon sequencing data produced via the short-read Illumina platform, but there are fewer options for long-read Oxford Nanopore Technologies (ONT) sequencing, or for processing data from environmental surveillance or other sources with many different organisms.
View Article and Find Full Text PDFJ Appl Microbiol
September 2025
Laboratory of Food Microbiology and Hygiene, Graduate School of Integrated Sciences for Life, Hiroshima University, 1-4-4 Kagamiyama, Higashihiroshima 739-8528, Japan.
Aims: This study aims to investigate the genomic profile of a multidrug-resistant Escherichia coli strain, 160-11H1, co-carrying an extended-spectrum β-lactamase (ESBL) and the plasmid-mediated mobile colistin resistance gene, mcr-5.
Methods And Results: The entire genome of the strain was sequenced using Illumina MiSeq and Oxford Nanopore platforms, and de novo assembly was performed using Unicycler. The genome size was 5 031,330 bp and comprised 5 140 coding sequences.
Microbiol Spectr
September 2025
United States Department of Agriculture, Agricultural Research Service (USDA-ARS), Southeast Poultry Research Laboratories, US National Poultry Research Center, Athens, Georgia, USA.
Infectious bursal disease (IBD), a highly contagious viral disease in young chickens, poses significant economic losses due to high mortality and immunosuppression. While IBD virus (IBDV) virulence is influenced by multiple genes, whole-genome sequencing (WGS) of IBDV is crucial for defining the strain pathotype and clinical profile. Flinders Technology Associates (FTA) cards are convenient for field sample collection, but their filter paper matrix can hinder nucleic acid recovery, impacting sequencing efficiency.
View Article and Find Full Text PDFMicrobiol Resour Announc
September 2025
Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA.
The complete genomes of three isolates were collected from the same patient in 2014, 2015, and 2016. NIH-016-2 is susceptible to eight antibiotics, while the other two are resistant to 11 commonly used antibiotics. Isolates were sequenced using Nanopore and Illumina short-read technologies.
View Article and Find Full Text PDFInt J Syst Evol Microbiol
September 2025
School of Biomolecular and Biomedical Science, Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland.
Two yeast strains, PYCC 10015 and PYCC 10016, were isolated from soil from an Irish forest. Sequence analysis of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) of the rRNA gene repeat, and the D1/D2 domain of the LSU rRNA gene, showed that they belong to the and genera of the order , but they did not exactly match any known species.
View Article and Find Full Text PDF