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Protocol for single-cell isolation and genome amplification of environmental microbial eukaryotes for genomic analysis
We describe environmental microbial eukaryotes (EMEs) sample collection, single-cell isolation, lysis, and genome amplification, followed by the rDNA amplification and OTU screening for recovery of high-quality species-specific genomes for de novo assembly. These protocols are part of our pipeline t...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8760549/ https://www.ncbi.nlm.nih.gov/pubmed/35059648 http://dx.doi.org/10.1016/j.xpro.2021.100968 |
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author | Ciobanu, Doina Chan, Sandy Ahrendt, Steven Quandt, C. Alisha Benny, Gerald L. Smith, Matthew E. James, Timothy Y. Cheng, Jan-Fang |
author_facet | Ciobanu, Doina Chan, Sandy Ahrendt, Steven Quandt, C. Alisha Benny, Gerald L. Smith, Matthew E. James, Timothy Y. Cheng, Jan-Fang |
author_sort | Ciobanu, Doina |
collection | PubMed |
description | We describe environmental microbial eukaryotes (EMEs) sample collection, single-cell isolation, lysis, and genome amplification, followed by the rDNA amplification and OTU screening for recovery of high-quality species-specific genomes for de novo assembly. These protocols are part of our pipeline that also includes bioinformatic methods. The pipeline and its application on a wide range of phyla of different sample complexity are described in our complementary paper. In addition, this protocol describes optimized lysis, genome amplification, and OTU screening steps of the pipeline. For complete details on the use and execution of this protocol, please refer to Ciobanu et al. (2021). |
format | Online Article Text |
id | pubmed-8760549 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-87605492022-01-19 Protocol for single-cell isolation and genome amplification of environmental microbial eukaryotes for genomic analysis Ciobanu, Doina Chan, Sandy Ahrendt, Steven Quandt, C. Alisha Benny, Gerald L. Smith, Matthew E. James, Timothy Y. Cheng, Jan-Fang STAR Protoc Protocol We describe environmental microbial eukaryotes (EMEs) sample collection, single-cell isolation, lysis, and genome amplification, followed by the rDNA amplification and OTU screening for recovery of high-quality species-specific genomes for de novo assembly. These protocols are part of our pipeline that also includes bioinformatic methods. The pipeline and its application on a wide range of phyla of different sample complexity are described in our complementary paper. In addition, this protocol describes optimized lysis, genome amplification, and OTU screening steps of the pipeline. For complete details on the use and execution of this protocol, please refer to Ciobanu et al. (2021). Elsevier 2022-01-13 /pmc/articles/PMC8760549/ /pubmed/35059648 http://dx.doi.org/10.1016/j.xpro.2021.100968 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Ciobanu, Doina Chan, Sandy Ahrendt, Steven Quandt, C. Alisha Benny, Gerald L. Smith, Matthew E. James, Timothy Y. Cheng, Jan-Fang Protocol for single-cell isolation and genome amplification of environmental microbial eukaryotes for genomic analysis |
title | Protocol for single-cell isolation and genome amplification of environmental microbial eukaryotes for genomic analysis |
title_full | Protocol for single-cell isolation and genome amplification of environmental microbial eukaryotes for genomic analysis |
title_fullStr | Protocol for single-cell isolation and genome amplification of environmental microbial eukaryotes for genomic analysis |
title_full_unstemmed | Protocol for single-cell isolation and genome amplification of environmental microbial eukaryotes for genomic analysis |
title_short | Protocol for single-cell isolation and genome amplification of environmental microbial eukaryotes for genomic analysis |
title_sort | protocol for single-cell isolation and genome amplification of environmental microbial eukaryotes for genomic analysis |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8760549/ https://www.ncbi.nlm.nih.gov/pubmed/35059648 http://dx.doi.org/10.1016/j.xpro.2021.100968 |
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