Nearly finished genomes produced using gel microdroplet culturing reveal substantial intraspecies genomic diversity within the human microbiome

التفاصيل البيبلوغرافية
العنوان: Nearly finished genomes produced using gel microdroplet culturing reveal substantial intraspecies genomic diversity within the human microbiome
المؤلفون: Karen W. Davenport, Cliff Han, Michael S. Fitzsimons, Jeremy P. Snook, Patti L. Wills, Armand E. K. Dichosa, Cheryl D. Gleasner, Krista G. Reitenga, Roger S. Lasken, Mark Novotny, Kim McMurry, Patrick S. G. Chain, John C. Detter, Shannon L. Johnson, Jian He, Wei Gu, Chien-Chi Lo, Ashlynn R. Daughton, B. Parson-Quintana, Joyclyn Yee-Greenbaum, Olga Chertkov
المصدر: Genome Research. 23:878-888
بيانات النشر: Cold Spring Harbor Laboratory, 2013.
سنة النشر: 2013
مصطلحات موضوعية: Genetics, Genome evolution, Bacteria, Microbiota, Multiple displacement amplification, Genetic Variation, Method, Genomics, Sequence Analysis, DNA, Bacterial genome size, Genome project, Biology, Polymerase Chain Reaction, Polymorphism, Single Nucleotide, Genome, DNA sequencing, Humans, Genome size, Genome, Bacterial, Genetics (clinical)
الوصف: The majority of microbial genomic diversity remains unexplored. This is largely due to our inability to culture most microorganisms in isolation, which is a prerequisite for traditional genome sequencing. Single-cell sequencing has allowed researchers to circumvent this limitation. DNA is amplified directly from a single cell using the whole-genome amplification technique of multiple displacement amplification (MDA). However, MDA from a single chromosome copy suffers from amplification bias and a large loss of specificity from even very small amounts of DNA contamination, which makes assembling a genome difficult and completely finishing a genome impossible except in extraordinary circumstances. Gel microdrop cultivation allows culturing of a diverse microbial community and provides hundreds to thousands of genetically identical cells as input for an MDA reaction. We demonstrate the utility of this approach by comparing sequencing results of gel microdroplets and single cells following MDA. Bias is reduced in the MDA reaction and genome sequencing, and assembly is greatly improved when using gel microdroplets. We acquired multiple near-complete genomes for two bacterial species from human oral and stool microbiome samples. A significant amount of genome diversity, including single nucleotide polymorphisms and genome recombination, is discovered. Gel microdroplets offer a powerful and high-throughput technology for assembling whole genomes from complex samples and for probing the pan-genome of naturally occurring populations.
تدمد: 1088-9051
DOI: 10.1101/gr.142208.112
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9f99dd375a215b9a8b416ac110301a49
https://doi.org/10.1101/gr.142208.112
Rights: OPEN
رقم الانضمام: edsair.doi.dedup.....9f99dd375a215b9a8b416ac110301a49
قاعدة البيانات: OpenAIRE
الوصف
تدمد:10889051
DOI:10.1101/gr.142208.112