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Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit

RNA and DNA extraction is a requirement for the study of gene expression and has an increasingly important role in genetic studies of all fleshy fruits. RNA and DNA extraction is difficult in kiwifruit due to the significant amount of polysaccharides and polyphenols compounds. So far, no commercial...

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Autores principales: Afshar-Mohammadian, Mansour, Rezadoost, Mohammad Hossein, Fallah, Seyyed Fatemeh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6058074/
https://www.ncbi.nlm.nih.gov/pubmed/30050755
http://dx.doi.org/10.1016/j.mex.2018.03.008
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author Afshar-Mohammadian, Mansour
Rezadoost, Mohammad Hossein
Fallah, Seyyed Fatemeh
author_facet Afshar-Mohammadian, Mansour
Rezadoost, Mohammad Hossein
Fallah, Seyyed Fatemeh
author_sort Afshar-Mohammadian, Mansour
collection PubMed
description RNA and DNA extraction is a requirement for the study of gene expression and has an increasingly important role in genetic studies of all fleshy fruits. RNA and DNA extraction is difficult in kiwifruit due to the significant amount of polysaccharides and polyphenols compounds. So far, no commercial kit has been developed specifically for high-quality RNA and DNA extraction in kiwifruit and the common protocols for RNA extraction have poor yields. This study developed a new protocol for high quality RNA extraction in Actinidia deliciosa. According to the results, the average yield of RNA extraction of fruit and leaf of A. deliciosa was ∼2180.7 ng/μl (∼545.175 μg/g FW) and ∼3424.9 ng/μl (∼856.225 μg/g FW), respectively with A(260)/A(280) between 1.95 to 2.07 and A(260)/A(230) higher than 2 indicating high RNA purity. While the averages yield of RNA extraction using previous methods from kiwifruit and leaf was 23 μg/g FW and 527 μg/g FW, respectively. Also, the average yields of genomic DNA from kiwifruit ranged from 52 to 98 ng/μl with A(260)/A(230) between 0.60 to 1.64 and A(260)/A(280) between 1.40 to 1.48. To our knowledge, this is the first report of a highly efficient and rapid method of RNA and DNA extraction in kiwifruit which can be used for a broad spectrum of the all fleshy fruits.
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spelling pubmed-60580742018-07-26 Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit Afshar-Mohammadian, Mansour Rezadoost, Mohammad Hossein Fallah, Seyyed Fatemeh MethodsX Biochemistry, Genetics and Molecular Biology RNA and DNA extraction is a requirement for the study of gene expression and has an increasingly important role in genetic studies of all fleshy fruits. RNA and DNA extraction is difficult in kiwifruit due to the significant amount of polysaccharides and polyphenols compounds. So far, no commercial kit has been developed specifically for high-quality RNA and DNA extraction in kiwifruit and the common protocols for RNA extraction have poor yields. This study developed a new protocol for high quality RNA extraction in Actinidia deliciosa. According to the results, the average yield of RNA extraction of fruit and leaf of A. deliciosa was ∼2180.7 ng/μl (∼545.175 μg/g FW) and ∼3424.9 ng/μl (∼856.225 μg/g FW), respectively with A(260)/A(280) between 1.95 to 2.07 and A(260)/A(230) higher than 2 indicating high RNA purity. While the averages yield of RNA extraction using previous methods from kiwifruit and leaf was 23 μg/g FW and 527 μg/g FW, respectively. Also, the average yields of genomic DNA from kiwifruit ranged from 52 to 98 ng/μl with A(260)/A(230) between 0.60 to 1.64 and A(260)/A(280) between 1.40 to 1.48. To our knowledge, this is the first report of a highly efficient and rapid method of RNA and DNA extraction in kiwifruit which can be used for a broad spectrum of the all fleshy fruits. Elsevier 2018-04-10 /pmc/articles/PMC6058074/ /pubmed/30050755 http://dx.doi.org/10.1016/j.mex.2018.03.008 Text en © 2018 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Biochemistry, Genetics and Molecular Biology
Afshar-Mohammadian, Mansour
Rezadoost, Mohammad Hossein
Fallah, Seyyed Fatemeh
Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit
title Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit
title_full Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit
title_fullStr Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit
title_full_unstemmed Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit
title_short Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit
title_sort comparative analysis and innovation of a simple and rapid method for high-quality rna and dna extraction of kiwifruit
topic Biochemistry, Genetics and Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6058074/
https://www.ncbi.nlm.nih.gov/pubmed/30050755
http://dx.doi.org/10.1016/j.mex.2018.03.008
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