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Updated: Oct 9, 2025

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
Published on: August 29, 2014
An accessible, efficient and global approach for the large-scale sequencing of bacterial genomes
Blanca M Perez-Sepulveda1, Darren Heavens2, Caisey V Pulford3
1Institute of Infection, Veterinary & Ecological Sciences, University of Liverpool, Liverpool, UK. blanca.m.perez@gmail.com.
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. Following the optimization of a logistics pipeline that involved shipping isolates as thermolysates in ambient conditions, the project assembled a diverse collection of 10,419 isolates from low- and middle-income countries. The genomes were sequenced using the LITE pipeline for library construction, with a total reagent cost of less than USD$10 per genome. Our method can be applied to other large bacterial collections to underpin global collaborations.
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