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Decontamination of MDA Reagents for Single Cell Whole Genome Amplification

Single cell genomics is a powerful and increasingly popular tool for studying the genetic make-up of uncultured microbes. A key challenge for successful single cell sequencing and analysis is the removal of exogenous DNA from whole genome amplification reagents. We found that UV irradiation of the m...

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Autores principales: Woyke, Tanja, Sczyrba, Alexander, Lee, Janey, Rinke, Christian, Tighe, Damon, Clingenpeel, Scott, Malmstrom, Rex, Stepanauskas, Ramunas, Cheng, Jan-Fang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3197606/
https://www.ncbi.nlm.nih.gov/pubmed/22028825
http://dx.doi.org/10.1371/journal.pone.0026161
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author Woyke, Tanja
Sczyrba, Alexander
Lee, Janey
Rinke, Christian
Tighe, Damon
Clingenpeel, Scott
Malmstrom, Rex
Stepanauskas, Ramunas
Cheng, Jan-Fang
author_facet Woyke, Tanja
Sczyrba, Alexander
Lee, Janey
Rinke, Christian
Tighe, Damon
Clingenpeel, Scott
Malmstrom, Rex
Stepanauskas, Ramunas
Cheng, Jan-Fang
author_sort Woyke, Tanja
collection PubMed
description Single cell genomics is a powerful and increasingly popular tool for studying the genetic make-up of uncultured microbes. A key challenge for successful single cell sequencing and analysis is the removal of exogenous DNA from whole genome amplification reagents. We found that UV irradiation of the multiple displacement amplification (MDA) reagents, including the Phi29 polymerase and random hexamer primers, effectively eliminates the amplification of contaminating DNA. The methodology is quick, simple, and highly effective, thus significantly improving whole genome amplification from single cells.
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spelling pubmed-31976062011-10-25 Decontamination of MDA Reagents for Single Cell Whole Genome Amplification Woyke, Tanja Sczyrba, Alexander Lee, Janey Rinke, Christian Tighe, Damon Clingenpeel, Scott Malmstrom, Rex Stepanauskas, Ramunas Cheng, Jan-Fang PLoS One Research Article Single cell genomics is a powerful and increasingly popular tool for studying the genetic make-up of uncultured microbes. A key challenge for successful single cell sequencing and analysis is the removal of exogenous DNA from whole genome amplification reagents. We found that UV irradiation of the multiple displacement amplification (MDA) reagents, including the Phi29 polymerase and random hexamer primers, effectively eliminates the amplification of contaminating DNA. The methodology is quick, simple, and highly effective, thus significantly improving whole genome amplification from single cells. Public Library of Science 2011-10-20 /pmc/articles/PMC3197606/ /pubmed/22028825 http://dx.doi.org/10.1371/journal.pone.0026161 Text en Woyke et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Woyke, Tanja
Sczyrba, Alexander
Lee, Janey
Rinke, Christian
Tighe, Damon
Clingenpeel, Scott
Malmstrom, Rex
Stepanauskas, Ramunas
Cheng, Jan-Fang
Decontamination of MDA Reagents for Single Cell Whole Genome Amplification
title Decontamination of MDA Reagents for Single Cell Whole Genome Amplification
title_full Decontamination of MDA Reagents for Single Cell Whole Genome Amplification
title_fullStr Decontamination of MDA Reagents for Single Cell Whole Genome Amplification
title_full_unstemmed Decontamination of MDA Reagents for Single Cell Whole Genome Amplification
title_short Decontamination of MDA Reagents for Single Cell Whole Genome Amplification
title_sort decontamination of mda reagents for single cell whole genome amplification
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3197606/
https://www.ncbi.nlm.nih.gov/pubmed/22028825
http://dx.doi.org/10.1371/journal.pone.0026161
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