title: RNASwift: A rapid, versatile RNA extraction method free from phenol and chloroform.
abstract: RNASwift is an inexpensive, versatile method for the rapid extraction of RNA. Existing RNA extraction methods typically use hazardous chemicals including phenol, chloroform and formamide which are often difficult to completely remove from the extracted RNA. RNASwift uses sodium chloride and sodium dodecyl sulphate to lyse the cells and isolate the RNA from the abundant cellular components in conjunction with solid phase extraction or isopropanol precipitation to rapidly purify the RNA. Moreover, the purified RNA is directly compatible with downstream analysis. Using spectrophotometry in conjunction with ion pair reverse phase chromatography to analyse the extracted RNA, we show that RNASwift extracts and purifies RNA of higher quality and purity in comparison to alternative RNA extraction methods. The RNASwift method yields approximately 25 μg of RNA from only 10(8)Escherichia coli cells. Furthermore, RNASwift is versatile; the same simple reagents can be used to rapidly extract RNA from a variety of different cells including bacterial, yeast and mammalian cells. In addition to the extraction of total RNA, the RNASwift method can also be used to extract double stranded RNA from genetically modified E. coli in higher yields compared to alternative methods.
authors: Department of Chemical and Biological Engineering, ChELSI Institute, Mappin Street, University of Sheffield, S1 3JD, UK.
authors: Alison O
authors: AO
authors: Nwokeoji
authors: Syngenta, Jealott's Hill International Research Centre, Bracknell, Berkshire, RG42 6EY, UK.
authors: Peter M
authors: PM
authors: Kilby
authors: Syngenta, Jealott's Hill International Research Centre, Bracknell, Berkshire, RG42 6EY, UK.
authors: David E
authors: DE
authors: Portwood
authors: Department of Chemical and Biological Engineering, ChELSI Institute, Mappin Street, University of Sheffield, S1 3JD, UK. Electronic address: m.dickman@sheffield.ac.uk.
authors: Mark J
authors: MJ
authors: Dickman
mesh_terms: D004926:Escherichia coli / Q000737:chemistry*; D012329:RNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D012329:RNA, Bacterial
keywords: Ion pair reverse phase HPLC; RNA extraction; RNA purification; UV spectrophotometry