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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...

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Detalles Bibliográficos
Autores principales: Ciobanu, Doina, Chan, Sandy, Ahrendt, Steven, Quandt, C. Alisha, Benny, Gerald L., Smith, Matthew E., James, Timothy Y., Cheng, Jan-Fang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
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).
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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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