Elsevier

Analytical Biochemistry

Volume 387, Issue 1, 1 April 2009, Pages 136-138
Analytical Biochemistry

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A simple and loss-free method to remove TRIzol contaminations from minute RNA samples

https://doi.org/10.1016/j.ab.2008.12.020Get rights and content

Abstract

Acid guanidium phenol preparations such as TRIzol allow the reproducible isolation of high-quality total RNA from various sources. However, if applied to minimal sample sizes, the quality parameters of the isolated RNA are often low. Here we present an approach to improve the 260/280- and 230/260-nm ratios of such preparations as well as the ratio of the 18S/28S RNA. A simple extraction with 1-butanol eliminates smearing of the 28 S RNA and restores the characteristic ultraviolet (UV) spectrum of highly purified RNA. Application of the method is demonstrated for fluorescence-activated cell sorting (FACS)-sorted cells where the population of interest is often small.

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Acknowledgments

The authors thank G. J. Arnold for critical review and helpful discussions during manuscript composition. The presented work was supported by the German Federal Ministry of Education and Research (BMBF, grant 0315128A, “Remedy”) within the framework of the FUGATO-plus program. The Deutsche Forschungsgemeinschaft (FOR 585) also supported this work.

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