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RNASwift:一种快速、通用的无酚和氯仿的RNA提取方法。

RNASwift: A rapid, versatile RNA extraction method free from phenol and chloroform.

作者信息

Nwokeoji Alison O, Kilby Peter M, Portwood David E, Dickman Mark J

机构信息

Department of Chemical and Biological Engineering, ChELSI Institute, Mappin Street, University of Sheffield, S1 3JD, UK.

Syngenta, Jealott's Hill International Research Centre, Bracknell, Berkshire, RG42 6EY, UK.

出版信息

Anal Biochem. 2016 Nov 1;512:36-46. doi: 10.1016/j.ab.2016.08.001. Epub 2016 Aug 3.

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.

摘要

RNASwift是一种用于快速提取RNA的低成本、多功能方法。现有的RNA提取方法通常使用包括苯酚、氯仿和甲酰胺在内的危险化学品,这些化学品往往难以从提取的RNA中完全去除。RNASwift使用氯化钠和十二烷基硫酸钠裂解细胞,并结合固相萃取或异丙醇沉淀从丰富的细胞成分中分离RNA,以快速纯化RNA。此外,纯化后的RNA可直接用于下游分析。通过使用分光光度法结合离子对反相色谱法分析提取的RNA,我们发现与其他RNA提取方法相比,RNASwift提取和纯化的RNA质量和纯度更高。RNASwift方法仅从10⁸个大肠杆菌细胞中就能产生约25μg的RNA。此外,RNASwift具有通用性;相同的简单试剂可用于从包括细菌、酵母和哺乳动物细胞在内的各种不同细胞中快速提取RNA。除了提取总RNA外,与其他方法相比,RNASwift方法还可用于从转基因大肠杆菌中以更高的产量提取双链RNA。

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