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An improved method for the rapid isolation of RNA from Arabidopsis and seeds of other species high in polyphenols and polysaccharides

Published online by Cambridge University Press:  07 August 2018

Steven Footitt ,
Sajjad Awan  and
William E. Finch-Savage
Steven Footitt*
Affiliation:
School of Life Sciences, Wellesbourne Campus, Warwick University, Wellesbourne, Warwick CV35 9EF, UK
Sajjad Awan
Affiliation:
School of Life Sciences, Wellesbourne Campus, Warwick University, Wellesbourne, Warwick CV35 9EF, UK
William E. Finch-Savage
Affiliation:
School of Life Sciences, Wellesbourne Campus, Warwick University, Wellesbourne, Warwick CV35 9EF, UK
*
Author for correspondence: Steven Footitt, Email: sfootitt@outlook.com
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Abstract

Seeds are notoriously high in polyphenols and polysaccharides, which reduce RNA quality and yield, and interfere with downstream applications. We present simple modifications to a rapid RNA extraction protocol for use with seeds. The inclusion of polyethylene glycol in place of polyvinylpyrrolidone reduced polyphenol and polysaccharide contamination. In addition, replacing NaCl with KCl improved the RNA yield from Arabidopsis seeds still bound by mucilage. On extraction of Arabidopsis seed recovered from field soils clean RNA pellets with no accompanying gelatinous matrix (polysaccharide) were seen, with A260/230 ratios greater than 1.8 confirming the lack of polysaccharide carry-over. When Brassica oleracea and Sinapis arvensis seeds were extracted, 260/230 ratios greater than 1.8 were seen. RNA yields in excess of 10 µg per 100 mg seed suitable for RT-QPCR were obtained.

Keywords

Arabidopsis thalianaBrassicapolysaccharideRNA extractionseeds

Information

Type
Technical Update
Information
Seed Science Research , Volume 28 , Issue 4 , December 2018 , pp. 360 - 364
Copyright
Copyright © Cambridge University Press 2018 

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