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• W2092551624 abstract "The study of bacterial populations using whole-genome sequencing is of considerable scientific and clinical interest. However, obtaining bacterial genomic information is not always trivial: the target bacteria may be difficult to culture or uncultured, and they may be found within samples containing complex mixtures of other contaminating microbes and/or host cells, from which it is very difficult to derive robust sequencing data. Here we describe our procedure to generate sufficient DNA for whole-genome sequencing from clinical samples and without the need for culture, as successfully used on the difficult-to-culture, obligate intracellular pathogen Chlamydia trachomatis. Our protocol combines immunomagnetic separation (IMS) for targeted bacterial enrichment with multiple displacement amplification (MDA) for whole-genome amplification (WGA), which is followed by high-throughput sequencing. Compared with other techniques that might be used to generate such data, IMS-MDA is an inexpensive, low-technology and highly transferable process that provides amplified genomic DNA for sequencing from target bacteria in under 5 h, with little hands-on time." @default.
• W2092551624 created "2016-06-24" @default.
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• W2092551624 date "2013-11-07" @default.
• W2092551624 modified "2023-10-13" @default.
• W2092551624 title "Generating whole bacterial genome sequences of low-abundance species from complex samples with IMS-MDA" @default.
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• W2092551624 doi "https://doi.org/10.1038/nprot.2013.147" @default.
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