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A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense
Acquiring high quality RNA is the basis of plant molecular biology research, plant genetics and other physiological investigations. At present, a large number of nucleotide isolation methods have been exploited or modified, such as commercial kits, CTAB, SDS methods and so on. Due to the nature of d...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5929529/ https://www.ncbi.nlm.nih.gov/pubmed/29715304 http://dx.doi.org/10.1371/journal.pone.0196592 |
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author | Liu, Lulu Han, Rongchun Yu, Nianjun Zhang, Wei Xing, Lihua Xie, Dongmei Peng, Daiyin |
author_facet | Liu, Lulu Han, Rongchun Yu, Nianjun Zhang, Wei Xing, Lihua Xie, Dongmei Peng, Daiyin |
author_sort | Liu, Lulu |
collection | PubMed |
description | Acquiring high quality RNA is the basis of plant molecular biology research, plant genetics and other physiological investigations. At present, a large number of nucleotide isolation methods have been exploited or modified, such as commercial kits, CTAB, SDS methods and so on. Due to the nature of different plants, extraction methods vary. Moreover, efficiency of certain approach cannot be guaranteed due to composition of different plants and extracting high quality RNA from plants rich in polysaccharides and polyphenols are often difficult. The physical and chemical properties of polysaccharides which are similar to nucleic acids and other secondary metabolites will be coprecipitated with RNA irreversibly. Therefore, how to remove polysaccharides and other secondary metabolites during RNA extraction is the primary challenge. Dendrobium huoshanense is an Orchidaceae perennial herb that is rich in polysaccharides and other secondary metabolites. By using D. huoshanense as the subject, we improved the method originated from CHAN and made it suitable for plants containing high amount of polysaccharides and polyphenols. The extracted total RNA was clear and non-dispersive, with good integrity and no obvious contamination with DNA and other impurities. And it was also evaluated by gel electrophoresis, nucleic acid quantitative detector and PCR assessment. Thus, as a simple approach, it is suitable and efficient in RNA isolation for plants rich in polysaccharides and polyphenols. |
format | Online Article Text |
id | pubmed-5929529 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-59295292018-05-11 A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense Liu, Lulu Han, Rongchun Yu, Nianjun Zhang, Wei Xing, Lihua Xie, Dongmei Peng, Daiyin PLoS One Research Article Acquiring high quality RNA is the basis of plant molecular biology research, plant genetics and other physiological investigations. At present, a large number of nucleotide isolation methods have been exploited or modified, such as commercial kits, CTAB, SDS methods and so on. Due to the nature of different plants, extraction methods vary. Moreover, efficiency of certain approach cannot be guaranteed due to composition of different plants and extracting high quality RNA from plants rich in polysaccharides and polyphenols are often difficult. The physical and chemical properties of polysaccharides which are similar to nucleic acids and other secondary metabolites will be coprecipitated with RNA irreversibly. Therefore, how to remove polysaccharides and other secondary metabolites during RNA extraction is the primary challenge. Dendrobium huoshanense is an Orchidaceae perennial herb that is rich in polysaccharides and other secondary metabolites. By using D. huoshanense as the subject, we improved the method originated from CHAN and made it suitable for plants containing high amount of polysaccharides and polyphenols. The extracted total RNA was clear and non-dispersive, with good integrity and no obvious contamination with DNA and other impurities. And it was also evaluated by gel electrophoresis, nucleic acid quantitative detector and PCR assessment. Thus, as a simple approach, it is suitable and efficient in RNA isolation for plants rich in polysaccharides and polyphenols. Public Library of Science 2018-05-01 /pmc/articles/PMC5929529/ /pubmed/29715304 http://dx.doi.org/10.1371/journal.pone.0196592 Text en © 2018 Liu 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 (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Liu, Lulu Han, Rongchun Yu, Nianjun Zhang, Wei Xing, Lihua Xie, Dongmei Peng, Daiyin A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense |
title | A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense |
title_full | A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense |
title_fullStr | A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense |
title_full_unstemmed | A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense |
title_short | A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense |
title_sort | method for extracting high-quality total rna from plant rich in polysaccharides and polyphenols using dendrobium huoshanense |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5929529/ https://www.ncbi.nlm.nih.gov/pubmed/29715304 http://dx.doi.org/10.1371/journal.pone.0196592 |
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