A monophasic solution for isolation of RNA devoid of polymerase chain reaction-detectable genomic DNA contamination

Ujjal Ghosh; Shankhamala Bose; Trinath Ghosh; Tapas Kumar Bandyopadhyay; Utpal Basu

doi:10.1016/j.ab.2015.02.018

A monophasic solution for isolation of RNA devoid of polymerase chain reaction-detectable genomic DNA contamination

Anal Biochem. 2015 May 15:477:50-2. doi: 10.1016/j.ab.2015.02.018. Epub 2015 Feb 27.

Authors

Ujjal Ghosh¹, Shankhamala Bose¹, Trinath Ghosh¹, Tapas Kumar Bandyopadhyay¹, Utpal Basu²

Affiliations

¹ Department of Molecular Biology & Biotechnology, University of Kalyani, Kalyani 741235, India.
² Department of Molecular Biology & Biotechnology, University of Kalyani, Kalyani 741235, India. Electronic address: utpal@klyuniv.ac.in.

PMID: 25731759
DOI: 10.1016/j.ab.2015.02.018

Abstract

Commercially available reagents and published protocols are widely used for RNA isolation. However, genomic DNA contamination in isolated RNA is a potential problem. Here we describe a simple, inexpensive method for eliminating genomic DNA contamination beyond the level of PCR-based detection through reduction of the guanidine thiocyanate concentration (1.5M) in a single monophasic solution based on Chomczynski-Sacchi reagents. The new method can be used to isolate small and large RNA species of high quality and can be completed within an hour.

Keywords: Monophasic solution; PCR; RNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
DNA Contamination*
Genomics*
Mice
Polymerase Chain Reaction
RNA / isolation & purification*

Substances

RNA