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GrainGenes Reference Report: PDS-79-246

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Reference	PDS-79-246
Title	Extraction of double-stranded RNA from plant tissues without the use of organic solvents
Journal	Plant Disease
Year	1995
Volume	79
Pages	246-248
Author	DePaulo J Powell C
Abstract	Double-stranded RNAs (dsRNAs) are commonly isolated from plants and insects with toxic organic solvents such as phenol and chloroform that denature and remove proteins from aqueous solutions containing nucleic acids This procedure was modified by replacing the toxic solvents with sodium dodecyl sulfate and potassium acetate (SDS/KOAc) Samples were homogenized in liquid nitrogen mixed with buffer and SDS at 65 C and then with KOAc at 0 C A precipitate, consisting of SDS, KOAc, and proteins, was removed by centrifugation. The supernatant containing nucleic acids was collected and subjected to standard CF-11 cellulose chromatography to separate dsRNAs from other nucleic acids. The SDS/KOAc method produced comparable yields and purity of dsRNA products compared to the phenol/chloroform method and is considered a convenient, effective alternative for isolating dsRNA from plant tissues.
Keyword	[ Hide all but 1 of 16 ] acetates-salts citrus citrus-excelsa citrus-tristeza-closterovirus extraction hordeum vulgare isolation-techniques leaves nicotiana tabacum phloem plant viruses plants potassium-acetate rna sodium-dodecylbenzenesulfonate tobacco-mosaic-tobamovirus

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