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An optimized isolation protocol yields high‐quality RNA from cassava tissues (Manihot esculenta Crantz)

We developed and modified a precise, rapid, and reproducible protocol isolating high‐quality RNA from tissues of multiple varieties of cassava plants (Manihot esculenta Crantz). The resulting method is suitable for use in mini, midi, and maxi preparations and rapidly achieves high total RNA yields (...

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Autores principales: Behnam, Babak, Bohorquez‐Chaux, Adriana, Castaneda‐Mendez, Oscar Fernando, Tsuji, Hiroyuki, Ishitani, Manabu, Becerra Lopez‐Lavalle, Luis Augusto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6443859/
https://www.ncbi.nlm.nih.gov/pubmed/30984554
http://dx.doi.org/10.1002/2211-5463.12561
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author Behnam, Babak
Bohorquez‐Chaux, Adriana
Castaneda‐Mendez, Oscar Fernando
Tsuji, Hiroyuki
Ishitani, Manabu
Becerra Lopez‐Lavalle, Luis Augusto
author_facet Behnam, Babak
Bohorquez‐Chaux, Adriana
Castaneda‐Mendez, Oscar Fernando
Tsuji, Hiroyuki
Ishitani, Manabu
Becerra Lopez‐Lavalle, Luis Augusto
author_sort Behnam, Babak
collection PubMed
description We developed and modified a precise, rapid, and reproducible protocol isolating high‐quality RNA from tissues of multiple varieties of cassava plants (Manihot esculenta Crantz). The resulting method is suitable for use in mini, midi, and maxi preparations and rapidly achieves high total RNA yields (170–600 μg·g(−1)) using low‐cost chemicals and consumables and with minimal contamination from polysaccharides, polyphenols, proteins, and other secondary metabolites. In particular, A (260) : A (280) ratios were > 2.0 for RNA from various tissues, and all of the present RNA samples yielded ribosomal integrity number values of greater than six. The resulting high purity and quality of isolated RNA will facilitate downstream applications (quantitative reverse transcriptase‐polymerase chain reaction or RNA sequencing) in cassava molecular breeding.
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spelling pubmed-64438592019-04-12 An optimized isolation protocol yields high‐quality RNA from cassava tissues (Manihot esculenta Crantz) Behnam, Babak Bohorquez‐Chaux, Adriana Castaneda‐Mendez, Oscar Fernando Tsuji, Hiroyuki Ishitani, Manabu Becerra Lopez‐Lavalle, Luis Augusto FEBS Open Bio Method We developed and modified a precise, rapid, and reproducible protocol isolating high‐quality RNA from tissues of multiple varieties of cassava plants (Manihot esculenta Crantz). The resulting method is suitable for use in mini, midi, and maxi preparations and rapidly achieves high total RNA yields (170–600 μg·g(−1)) using low‐cost chemicals and consumables and with minimal contamination from polysaccharides, polyphenols, proteins, and other secondary metabolites. In particular, A (260) : A (280) ratios were > 2.0 for RNA from various tissues, and all of the present RNA samples yielded ribosomal integrity number values of greater than six. The resulting high purity and quality of isolated RNA will facilitate downstream applications (quantitative reverse transcriptase‐polymerase chain reaction or RNA sequencing) in cassava molecular breeding. John Wiley and Sons Inc. 2019-02-20 /pmc/articles/PMC6443859/ /pubmed/30984554 http://dx.doi.org/10.1002/2211-5463.12561 Text en © 2018 The Authors. Published by FEBS Press and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Method
Behnam, Babak
Bohorquez‐Chaux, Adriana
Castaneda‐Mendez, Oscar Fernando
Tsuji, Hiroyuki
Ishitani, Manabu
Becerra Lopez‐Lavalle, Luis Augusto
An optimized isolation protocol yields high‐quality RNA from cassava tissues (Manihot esculenta Crantz)
title An optimized isolation protocol yields high‐quality RNA from cassava tissues (Manihot esculenta Crantz)
title_full An optimized isolation protocol yields high‐quality RNA from cassava tissues (Manihot esculenta Crantz)
title_fullStr An optimized isolation protocol yields high‐quality RNA from cassava tissues (Manihot esculenta Crantz)
title_full_unstemmed An optimized isolation protocol yields high‐quality RNA from cassava tissues (Manihot esculenta Crantz)
title_short An optimized isolation protocol yields high‐quality RNA from cassava tissues (Manihot esculenta Crantz)
title_sort optimized isolation protocol yields high‐quality rna from cassava tissues (manihot esculenta crantz)
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6443859/
https://www.ncbi.nlm.nih.gov/pubmed/30984554
http://dx.doi.org/10.1002/2211-5463.12561
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