Efficient method for isolation of high-quality RNA from Psidium guajava L. tissues.

Acquiring high-quality RNA in sufficient amounts is crucial in plant molecular biology and genetic studies. Several methods for RNA extraction from plants are available in the literature, mainly due to the great biochemical diversity present in each species and tissue, which can complicate or preven...

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Autores principales:	Paola de Avelar Carpinetti, Vinicius Sartori Fioresi, Thais Ignez da Cruz, Francine Alves Nogueira de Almeida, Drielli Canal, Adésio Ferreira, Marcia Flores da Silva Ferreira
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b955e9a4765e4cb49c409f36b87fdf4a
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spelling	oai:doaj.org-article:b955e9a4765e4cb49c409f36b87fdf4a2021-12-02T20:06:25ZEfficient method for isolation of high-quality RNA from Psidium guajava L. tissues.1932-620310.1371/journal.pone.0255245https://doaj.org/article/b955e9a4765e4cb49c409f36b87fdf4a2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0255245https://doaj.org/toc/1932-6203Acquiring high-quality RNA in sufficient amounts is crucial in plant molecular biology and genetic studies. Several methods for RNA extraction from plants are available in the literature, mainly due to the great biochemical diversity present in each species and tissue, which can complicate or prevent the extraction. Psidium guajava (Myrtaceae family) is a perennial fruit tree of medicinal and economic value; nevertheless, only a few molecular studies are available for the species. One reason is the difficulty in obtaining RNA due to the content of the samples, which are rich in polyphenols, polysaccharides, and secondary metabolites. Furthermore, there are few studies available for the isolation of RNA from guava or Psidium samples, which hampers advances in the study of the genus. Here, quality and yields of RNA isolates were compared using six extraction protocols: two protocols based on the application of cetyltrimethylammonium bromide (CTAB) lysis buffer, one protocol which uses the TRIzol reagent, one which applies guanidine thiocyanate lysis buffer followed by organic phase extraction, and two commercial kits (PureLink RNA Mini Kit and RNeasy Plant Mini Kit). The CTAB-based method provided the highest RNA yields and quality for five different tissues (flower bud, immature leaf, young leaf, mature leaf, and root), genotypes, and stress conditions. For the most efficient protocol, the average yield of RNA from guava leaves was 203.06 μg/g of tissue, and the A260/A280 and A260/A230 ratios were 2.1 and 2.2, respectively. RT-qPCR analysis demonstrated that the purity of the samples was sufficient for molecular biology experiments. CTAB-based methods for RNA isolation were found to be the most efficient, providing the highest RNA yields and quality for tissues from P. guajava. Additionally, they were compatible for downstream RNA-based applications, besides being simple and cost-effective.Paola de Avelar CarpinettiVinicius Sartori FioresiThais Ignez da CruzFrancine Alves Nogueira de AlmeidaDrielli CanalAdésio FerreiraMarcia Flores da Silva FerreiraPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 7, p e0255245 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Paola de Avelar Carpinetti Vinicius Sartori Fioresi Thais Ignez da Cruz Francine Alves Nogueira de Almeida Drielli Canal Adésio Ferreira Marcia Flores da Silva Ferreira Efficient method for isolation of high-quality RNA from Psidium guajava L. tissues.
description	Acquiring high-quality RNA in sufficient amounts is crucial in plant molecular biology and genetic studies. Several methods for RNA extraction from plants are available in the literature, mainly due to the great biochemical diversity present in each species and tissue, which can complicate or prevent the extraction. Psidium guajava (Myrtaceae family) is a perennial fruit tree of medicinal and economic value; nevertheless, only a few molecular studies are available for the species. One reason is the difficulty in obtaining RNA due to the content of the samples, which are rich in polyphenols, polysaccharides, and secondary metabolites. Furthermore, there are few studies available for the isolation of RNA from guava or Psidium samples, which hampers advances in the study of the genus. Here, quality and yields of RNA isolates were compared using six extraction protocols: two protocols based on the application of cetyltrimethylammonium bromide (CTAB) lysis buffer, one protocol which uses the TRIzol reagent, one which applies guanidine thiocyanate lysis buffer followed by organic phase extraction, and two commercial kits (PureLink RNA Mini Kit and RNeasy Plant Mini Kit). The CTAB-based method provided the highest RNA yields and quality for five different tissues (flower bud, immature leaf, young leaf, mature leaf, and root), genotypes, and stress conditions. For the most efficient protocol, the average yield of RNA from guava leaves was 203.06 μg/g of tissue, and the A260/A280 and A260/A230 ratios were 2.1 and 2.2, respectively. RT-qPCR analysis demonstrated that the purity of the samples was sufficient for molecular biology experiments. CTAB-based methods for RNA isolation were found to be the most efficient, providing the highest RNA yields and quality for tissues from P. guajava. Additionally, they were compatible for downstream RNA-based applications, besides being simple and cost-effective.
format	article
author	Paola de Avelar Carpinetti Vinicius Sartori Fioresi Thais Ignez da Cruz Francine Alves Nogueira de Almeida Drielli Canal Adésio Ferreira Marcia Flores da Silva Ferreira
author_facet	Paola de Avelar Carpinetti Vinicius Sartori Fioresi Thais Ignez da Cruz Francine Alves Nogueira de Almeida Drielli Canal Adésio Ferreira Marcia Flores da Silva Ferreira
author_sort	Paola de Avelar Carpinetti
title	Efficient method for isolation of high-quality RNA from Psidium guajava L. tissues.
title_short	Efficient method for isolation of high-quality RNA from Psidium guajava L. tissues.
title_full	Efficient method for isolation of high-quality RNA from Psidium guajava L. tissues.
title_fullStr	Efficient method for isolation of high-quality RNA from Psidium guajava L. tissues.
title_full_unstemmed	Efficient method for isolation of high-quality RNA from Psidium guajava L. tissues.
title_sort	efficient method for isolation of high-quality rna from psidium guajava l. tissues.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/b955e9a4765e4cb49c409f36b87fdf4a
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Efficient method for isolation of high-quality RNA from Psidium guajava L. tissues.

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