論文 - 詳細
| RRC ID | 75594 |
|---|---|
| 著者 | Aoki H, Masahiro Y, Shimizu M, Hongoh Y, Ohkuma M, Yamagata Y. |
| タイトル | Agarose gel microcapsules enable easy-to-prepare, picolitre-scale, single-cell genomics, yielding high-coverage genome sequences. |
| ジャーナル | Sci Rep |
| Abstract |
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. |
| 巻・号 | 12(1) |
| ページ | 17014 |
| 公開日 | 2022-10-18 |
| DOI | 10.1038/s41598-022-20923-z |
| PII | 10.1038/s41598-022-20923-z |
| PMID | 36257967 |
| PMC | PMC9579161 |
| MeSH | Alginates Bacteria* / genetics Capsules Cytosine DNA Emulsions Genome, Bacterial Genomics* / methods Guanine Humans Sepharose Water |
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