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      A Rapid and Efficient Method for Purifying High Quality Total RNA from Peaches (Prunus persica) for Functional Genomics Analyses

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          Abstract

          Prunus persica has been proposed as a genomic model for deciduous trees and the Rosaceae family. Optimized protocols for RNA isolation are necessary to further advance studies in this model species such that functional genomics analyses may be performed. Here we present an optimized protocol to rapidly and efficiently purify high quality total RNA from peach fruits (Prunus persica). Isolating high-quality RNA from fruit tissue is often difficult due to large quantities of polysaccharides and polyphenolic compounds that accumulate in this tissue and co-purify with the RNA. Here we demonstrate that a modified version of the method used to isolate RNA from pine trees and the woody plant Cinnamomun tenuipilum is ideal for isolating high quality RNA from the fruits of Prunus persica. This RNA may be used for many functional genomic based experiments such as RT-PCR and the construction of large-insert cDNA libraries.

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          Most cited references19

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction

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            RNA isolation from highly viscous samples rich in polyphenols and polysaccharides

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              Improved method for the isolation of RNA from plant tissues.

              A fast and efficient method for the isolation of RNA from plant tissues is described. Tuber tissue is homogenized in a guanidine hydrochloride-containing buffer followed by direct extraction with phenol/chloroform. The RNA is precipitated from the aqueous phase, washed with 3 M sodium acetate and 70% ethanol, and finally dissolved in water. The yield of RNA is up to 500 micrograms/g of tissue and several tests indicate intact and nondegraded RNA. This method can be adapted to a small-scale version by the use of 1.5-ml tubes, allowing rapid isolation of RNA from a larger number of samples. Finally, this method is of particular use for isolating RNA from tissues with a high polysaccharide and nuclease content such as wounded potato tubers.
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                Author and article information

                Journal
                bres
                Biological Research
                Biol. Res.
                Sociedad de Biología de Chile (Santiago, , Chile )
                0716-9760
                2005
                : 38
                : 1
                : 83-88
                Affiliations
                [06] Santiago orgnameINIA La Platina Chile
                [03] Santiago orgnameUniversidad Andrés Bello orgdiv1Facultad de Ecología y Recursos Naturales orgdiv2Centro de Biotecnología Vegetal Chile
                [07] Santiago orgnameUniversidad de Chile orgdiv1Facultad de Ciencias Agronómicas Chile
                [01] orgnameMillenium Nucleus in Plant Cell Biology P02-009-F Chile
                [02] orgnameUniversidad de Chile orgdiv1Facultad de Ciencias orgdiv2Departamento de Biología Chile
                [04] Santiago orgnameUniversidad de Chile orgdiv1Facultad de Ciencias Físicas y Matemáticas Chile
                [05] Santiago orgnameUniversidad de Chile orgdiv1INTA Chile
                Article
                S0716-97602005000100010 S0716-9760(05)03800110
                10.4067/S0716-97602005000100010
                fe89e2ea-88ab-45ae-8887-d6e2f47fcb57

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 03 January 2005
                : 31 January 2005
                : 03 February 2005
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 20, Pages: 6
                Product

                SciELO Chile

                Categories
                Articles

                cDNA library,RT-PCR,Rosaceae,RNA isolation,Prunus persica,polysaccharides,peach,functional genomics,fruit trees,fruit

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