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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2011
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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. |
format | Online Article Text |
id | pubmed-3197606 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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