The DOI matches its Crossref record; 15 of the 15 references it cites were found in a registry or an index.
Nanoliter Reactors Improve Multiple Displacement Amplification of Genomes from Single Cells
Open accessAbstract
Since only a small fraction of environmental bacteria are amenable to laboratory culture, there is great interest in genomic sequencing directly from single cells. Sufficient DNA for sequencing can be obtained from one cell by the Multiple Displacement Amplification (MDA) method, thereby eliminating the need to develop culture methods. Here we used a microfluidic device to isolate individual Escherichia coli and amplify genomic DNA by MDA in 60-nl reactions. Our results confirm a report that reduced MDA reaction volume lowers nonspecific synthesis that can result from contaminant DNA templates and unfavourable interaction between primers. The quality of the genome amplification was assessed by qPCR and compared favourably to single-cell amplifications performed in standard 50-microl volumes. Amplification bias was greatly reduced in nanoliter volumes, thereby providing a more even representation of all sequences. Single-cell amplicons from both microliter and nanoliter volumes provided high-quality sequence data by high-throughput pyrosequencing, thereby demonstrating a straightforward route to sequencing genomes from single cells.
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Cites
Environmental genome shotgun sequencing of the sargasso sea
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Self-citationMicrofluidic Large-Scale Integration
Thorsen, T., Maerkl, S. J., & Quake, S. R. (2002) · Science · doi:10.1126/science.1076996 · Cited by 2,222
Self-citationThe Uncultured Microbial Majority
Rappé, M. S., & Giovannoni, S. J. (2003) · Annual Review of Microbiology · doi:10.1146/annurev.micro.57.030502.090759 · Cited by 2,017
Comprehensive human genome amplification using multiple displacement amplification
Dean, F. B., Hosono, S., Fang, L., Wu, X., Faruqi, A. F., Bray-Ward, P., Sun, Z., Zong, Q., Du, Y., Du, J., et al. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.082089499 · Cited by 1,467
Self-citationDean, F. B., Nelson, J. R., Giesler, T. L., & Lasken, R. S. (2001) · Genome Research · doi:10.1101/gr.180501 · Cited by 1,067
Self-citationMarcy, Y., Ouverney, C., Bik, E. M., Lösekann, T., Ivanova, N., Martin, H. G., Szeto, E., Platt, D., Hugenholtz, P., Relman, D. A., et al. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0704662104 · Cited by 634
Self-citationUnbiased Whole-Genome Amplification Directly From Clinical Samples
Hosono, S., Faruqi, A. F., Dean, F. B., Du, Y., Sun, Z., Wu, X., Du, J., Kingsmore, S. F., Egholm, M., & Lasken, R. S. (2003) · Genome Research · doi:10.1101/gr.816903 · Cited by 410
Self-citationSequencing genomes from single cells by polymerase cloning
Zhang, K., Martiny, A. C., Reppas, N. B., Barry, K. W., Malek, J., Chisholm, S. W., & Church, G. M. (2006) · Nature Biotechnology · doi:10.1038/nbt1214 · Cited by 410
Mechanism of chimera formation during the Multiple Displacement Amplification reaction
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Cited by
Single-cell genome sequencing: current state of the science
Gawad, C., Koh, W., & Quake, S. R. (2016) · Nature Reviews Genetics · doi:10.1038/nrg.2015.16 · Cited by 1,334
Self-citationDroplet microfluidics for high-throughput biological assays
Guo, M. T., Rotem, A., Heyman, J. A., & Weitz, D. A. (2012) · Lab on a Chip · doi:10.1039/c2lc21147e · Cited by 1,040
Quantitative assessment of single-cell RNA-sequencing methods
Wu, A. R., Neff, N. F., Kalisky, T., Dalerba, P., Treutlein, B., Rothenberg, M. E., Mburu, F. M., Mantalas, G. L., Sim, S., Clarke, M. F., et al. (2014) · Nature Methods · doi:10.1038/nmeth.2694 · Cited by 696
Self-citationGenome-wide Single-Cell Analysis of Recombination Activity and De Novo Mutation Rates in Human Sperm
Wang, J., Fan, H. C., Behr, B., & Quake, S. R. (2012) · Cell · doi:10.1016/j.cell.2012.06.030 · Cited by 511
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Metadata
- Authors
- Marcy, Y., Ishoey, T., Lasken, R. S., Stockwell, T. B., Walenz, B. P., Halpern, A. L., Beeson, K. Y., Goldberg, S. M. D., & Quake, S. R.
- Venue
- PLoS Genetics
- Publisher
- Public Library of Science (PLoS)
- Published
- 2007-09-21
- Volume
- 3
- Issue
- 9
- Pages
- e155
- ISSN
- 1553-7390 (ISSN-L)
- 1553-7404
- OpenAlex
- W2117273677
- Cited by
- 286
- References
- 15
- FWCI
- 5.75
- Type
- journal article
- Language
- en
- Open access
- gold
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- Marcy, Y., Ishoey, T., Lasken, R. S., Stockwell, T. B., Walenz, B. P., Halpern, A. L., Beeson, K. Y., Goldberg, S. M. D., & Quake, S. R. (2007). Nanoliter Reactors Improve Multiple Displacement Amplification of Genomes from Single Cells. PLoS Genetics. https://doi.org/10.1371/journal.pgen.0030155
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