Genome BiologyMicrobiologyPeer-reviewed journal article
An accessible, efficient and global approach for the large-scale sequencing of bacterial genomes
B. M. Perez-Sepulveda, D. Heavens, C. V. Pulford, A. V. Predeus, R. Low, H. Webster, C. Schudoma, W. Rowe, J. Lipscombe, C. Watkins, B. Kumwenda, N. Shearer, K. Costigan, K. S. Baker, N. A. Feasey, J. C. D. Hinton, N. Hall, The 10KSG consortium
Abstract
We have developed an efficient and inexpensive pipeline for streamlining large-scale collection and genome sequencing of bacterial isolates. Evaluation of this method involved a worldwide research collaboration focused on the model organism Salmonella enterica, the 10KSG consortium. By optimising a logistics pipeline that collected isolates as thermolysates, permitting shipment in ambient conditions, the project assembled a diverse collection of 10,419 clinical and environmental isolates from low- and middle-income countries in less than one year. The bacteria were obtained from 51 countries/territories dating from 1949 to 2017, with a focus on Africa and Latin-America. All isolates were collected in barcoded tubes and genome sequenced using an optimised DNA extraction method and the LITE pipeline for library construction. After Illumina sequencing, the total reagent cost was less than USD$10 per genome. Our method can be applied to genome-sequence other large bacterial collections at a relatively low cost, within a limited timeframe, to support global collaborations.
- organism: bacteria (other)
- assay: whole-genome sequencing
- research focus: variant calling
- approach: experimental assay
Reproducibility ledger
Nobody has reported trying to reproduce this paper yet. Did you replicate an experiment or run the code? Your report saves the next lab a week.
Sign in to file a reproducibility report.
Related papers
- Revolutionising Public Health Reference Microbiology using Whole Genome Sequencing: Salmonella as an exemplar
Philip Ashton, Satheesh Nair, … Kathie Grant·bioRxiv·Nov 29, 2015
- Low-cost genomics enable high-throughput isolate screening and strain-level microbiome profiling
J. G. Sanders, W. Yan, A. H. Moeller·bioRxiv·Apr 11, 2022
- Scalable Genotyping of Microbial Colonies
A. Chen, N. Nchinda, N. J. Cira·bioRxiv·Dec 6, 2023
- Inexpensive Multiplexed Library Preparation for Megabase-Sized Genomes
Michael Baym, Sergey Kryazhimskiy, … Roy Kishony·bioRxiv·Jan 21, 2015
- An optimised method for bacterial nucleic acid extraction from positive blood culture broths for whole genome sequencing, resistance phenotype prediction and downstream molecular applications
M. Bauer, A. M. Peri, … P. N. A. Harris·bioRxiv·Jun 30, 2022
- A mycobacterial DNA extraction protocol designed for resource limited settings generates high quality whole genome sequencing
Caitlin Percy, Ilinca Memelis, … Daire M. Cantillon·bioRxiv·May 31, 2024