A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants

Gambino, Giorgio; Perrone, Irene; Gribaudo, Ivana

doi:10.1002/pca.1078

Cited by 580 publications

(373 citation statements)

References 24 publications

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“…Although the extraction kits performed well for leaf and shoot tissues, clogging of columns was very common in extraction of tuber tissue. Slightly modified CTAB methods have been found useful for RNA extraction from many plant taxa that are rich in secondary metabolite content, e.g., Pinus (Chang et al 1993), Vitis (Gambino et al 2008), Hippophae (Ghangal et al 2009), and Hylocereus (Wong et al 2014). The protocol reported by us was modified mainly from the CTAB protocols of Chang et al (1993) and Ghangal et al (2009) as described below.…”

Section: Rna Extraction Methodologymentioning

confidence: 99%

A modified protocol yields high-quality RNA from highly mucilaginous Dioscorea tubers

2017

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Extraction of RNA from Dioscorea is difficult because of rich mucilage and secondary metabolite content. High-quality RNA is required for RT-PCR and transcriptome analysis. Different protocols and common extraction kits were used for RNA extraction in Dioscorea, but the results were not satisfactory. A CTAB based protocol with lithium chloride precipitation yielded good quality RNA. The RNA isolated using this protocol was successfully used for RT-PCR and transcriptome sequencing experiments. Mucilage content varies at different developmental stages of Dioscorea and the present protocol was effective in isolating RNA from such samples. Although the protocol was originally modified for tuber tissues, it can be used also for extraction of RNA from rhizome, root, shoot and leaf.

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Section: Rna Extraction Methodologymentioning

confidence: 99%

A modified protocol yields high-quality RNA from highly mucilaginous Dioscorea tubers

2017

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“…This is especially true for plants under abiotic and biotic stresses because of high accumulation of secondary metabolites (Winkel-Shirley 2002). The polysaccharides tend to co-precipitate with the RNA in the presence of alcohols, a major source of contamination in the final extract, and thus interfere with subsequent applications such as reverse transcription and cDNA library construction (Gambino et al 2008). While several commercial kits are available for successful extraction of RNA from many tissues, they have proved to be less effective on tissues rich in polyphenols and polysaccharides (Kiefer et al 2000).…”

Section: Introductionmentioning

confidence: 99%

Isolation of high quality RNA from pistachio (Pistacia vera L.) and other woody plants high in secondary metabolites

Jazi

Rajaei²,

Seyedi

2015

Physiol Mol Biol Plants

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The quality and quantity of RNA are critical for successful downstream transcriptome-based studies such as microarrays and RNA sequencing (RNA-Seq). RNA isolation from woody plants, such as Pistacia vera, with very high amounts of polyphenols and polysaccharides is an enormous challenge. Here, we describe a highly efficient protocol that overcomes the limitations posed by poor quality and low yield of isolated RNA from pistachio and various recalcitrant woody plants. The key factors that resulted in a yield of 150 μg of high quality RNA per 200 mg of plant tissue include the elimination of phenol from the extraction buffer, raising the concentration of β-mercaptoethanol, long time incubation at 65°C, and nucleic acid precipitation with optimized volume of NaCl and isopropyl alcohol. Also, the A260/A280 and A260/A230 of extracted RNA were about 1.9-2.1and 2.2-2.3, respectively, revealing the high purity. Since the isolated RNA passed highly stringent quality control standards for sensitive reactions, including RNA sequencing and real-time PCR, it can be considered as a reliable and cost-effective method for RNA extraction from woody plants.

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“…The difficulties of RNA extraction from plant tissues that are rich in polysaccharides such as grapevine, mango and sweet potatoes have been reported in many previous studies (Gambino et al 2008;Kim & Hamada 2005;López-Gómez & Gómez-Lim 1992). Different conditions/ parameters are therefore required to isolate a good quality RNA for different species, different tissues as well as for the same species but grown under different environments (Soon et al 1997).…”

Section: Introductionmentioning

confidence: 99%

RNA Extractions of Mangosteen (Garcinia mangostana L.) Pericarps for Sequencing

Abdul-Rahman

Suleman

Zakaria

et al. 2017

JSM

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A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants

Abstract: The study has shown that the improvement of a CTAB-based protocol allows the rapid isolation of high-quality RNA from grapevine and many woody species.

Cited by 580 publications

References 24 publications

A modified protocol yields high-quality RNA from highly mucilaginous Dioscorea tubers

A modified protocol yields high-quality RNA from highly mucilaginous Dioscorea tubers

Isolation of high quality RNA from pistachio (Pistacia vera L.) and other woody plants high in secondary metabolites

RNA Extractions of Mangosteen (Garcinia mangostana L.) Pericarps for Sequencing

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