期刊
SCIENTIFIC REPORTS
卷 12, 期 1, 页码 -出版社
NATURE PORTFOLIO
DOI: 10.1038/s41598-022-20923-z
关键词
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资金
- JSPS KAKENHI [16K07224, 17H01447, 19H05679]
- Institute of Fermentation, Osaka
- RIKEN Pioneering Project Biology of Symbiosis
A novel type of agarose gel microcapsule (AGM) has been developed to obtain high-quality, single-cell, amplified bacterial genomic DNA. The method is easy to prepare and does not require specialized equipment, allowing for easy and effective single-cell genomics and accelerated genomic analysis of yet-uncultured microorganisms.
A novel type of agarose gel microcapsule (AGM), consisting of an alginate picolitre sol core and an agarose gel shell, was developed to obtain high-quality, single-cell, amplified genomic DNA of bacteria. The AGM is easy to prepare in a stable emulsion with oil of water-equivalent density, which prevents AGM aggregation, with only standard laboratory equipment. Single cells from a pure culture of Escherichia coli, a mock community comprising 15 strains of human gut bacteria, and a termite gut bacterial community were encapsulated within AGMs, and their genomic DNA samples were prepared with massively parallel amplifications in a tube. The genome sequencing did not need second-round amplification and showed an average genome completeness that was much higher than that obtained using a conventional amplification method on the microlitre scale, regardless of the genomic guanine-cytosine content. Our novel method using AGM will allow many researchers to perform single-cell genomics easily and effectively, and can accelerate genomic analysis of yet-uncultured microorganisms.
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