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High quality RNA extraction from Maqui berry for its application in next-generation sequencing

Maqui berry (Aristotelia chilensis) is a native Chilean species that produces berries that are exceptionally rich in anthocyanins and natural antioxidants. These natural compounds provide an array of health benefits for humans, making them very desirable in a fruit. At the same time, these substance...

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Autores principales: Sánchez, Carolina, Villacreses, Javier, Blanc, Noelle, Espinoza, Loreto, Martinez, Camila, Pastor, Gabriela, Manque, Patricio, Undurraga, Soledad F., Polanco, Victor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4970997/
https://www.ncbi.nlm.nih.gov/pubmed/27536526
http://dx.doi.org/10.1186/s40064-016-2906-x
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author Sánchez, Carolina
Villacreses, Javier
Blanc, Noelle
Espinoza, Loreto
Martinez, Camila
Pastor, Gabriela
Manque, Patricio
Undurraga, Soledad F.
Polanco, Victor
author_facet Sánchez, Carolina
Villacreses, Javier
Blanc, Noelle
Espinoza, Loreto
Martinez, Camila
Pastor, Gabriela
Manque, Patricio
Undurraga, Soledad F.
Polanco, Victor
author_sort Sánchez, Carolina
collection PubMed
description Maqui berry (Aristotelia chilensis) is a native Chilean species that produces berries that are exceptionally rich in anthocyanins and natural antioxidants. These natural compounds provide an array of health benefits for humans, making them very desirable in a fruit. At the same time, these substances also interfere with nucleic acid preparations, making RNA extraction from Maqui berry a major challenge. Our group established a method for RNA extraction of Maqui berry with a high quality RNA (good purity, good integrity and higher yield). This procedure is based on the adapted CTAB method using high concentrations of PVP (4 %) and β-mercaptoethanol (4 %) and spermidine in the extraction buffer. These reagents help to remove contaminants such as polysaccharides, proteins, phenols and also prevent the oxidation of phenolic compounds. The high quality of RNA isolated through this method allowed its uses with success in molecular applications for this endemic Chilean fruit, such as differential expression analysis of RNA-Seq data using next generation sequencing (NGS). Furthermore, we consider that our method could potentially be used for other plant species with extremely high levels of antioxidants and anthocyanins.
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spelling pubmed-49709972016-08-17 High quality RNA extraction from Maqui berry for its application in next-generation sequencing Sánchez, Carolina Villacreses, Javier Blanc, Noelle Espinoza, Loreto Martinez, Camila Pastor, Gabriela Manque, Patricio Undurraga, Soledad F. Polanco, Victor Springerplus Methodology Maqui berry (Aristotelia chilensis) is a native Chilean species that produces berries that are exceptionally rich in anthocyanins and natural antioxidants. These natural compounds provide an array of health benefits for humans, making them very desirable in a fruit. At the same time, these substances also interfere with nucleic acid preparations, making RNA extraction from Maqui berry a major challenge. Our group established a method for RNA extraction of Maqui berry with a high quality RNA (good purity, good integrity and higher yield). This procedure is based on the adapted CTAB method using high concentrations of PVP (4 %) and β-mercaptoethanol (4 %) and spermidine in the extraction buffer. These reagents help to remove contaminants such as polysaccharides, proteins, phenols and also prevent the oxidation of phenolic compounds. The high quality of RNA isolated through this method allowed its uses with success in molecular applications for this endemic Chilean fruit, such as differential expression analysis of RNA-Seq data using next generation sequencing (NGS). Furthermore, we consider that our method could potentially be used for other plant species with extremely high levels of antioxidants and anthocyanins. Springer International Publishing 2016-08-03 /pmc/articles/PMC4970997/ /pubmed/27536526 http://dx.doi.org/10.1186/s40064-016-2906-x Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Methodology
Sánchez, Carolina
Villacreses, Javier
Blanc, Noelle
Espinoza, Loreto
Martinez, Camila
Pastor, Gabriela
Manque, Patricio
Undurraga, Soledad F.
Polanco, Victor
High quality RNA extraction from Maqui berry for its application in next-generation sequencing
title High quality RNA extraction from Maqui berry for its application in next-generation sequencing
title_full High quality RNA extraction from Maqui berry for its application in next-generation sequencing
title_fullStr High quality RNA extraction from Maqui berry for its application in next-generation sequencing
title_full_unstemmed High quality RNA extraction from Maqui berry for its application in next-generation sequencing
title_short High quality RNA extraction from Maqui berry for its application in next-generation sequencing
title_sort high quality rna extraction from maqui berry for its application in next-generation sequencing
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4970997/
https://www.ncbi.nlm.nih.gov/pubmed/27536526
http://dx.doi.org/10.1186/s40064-016-2906-x
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