Authors: Ince, Ayşe Gül; Karaca, Mehmet

Source: Journal of the Science of Food and Agriculture, Volume 89, Number 1, 15 January 2009 , pp. 168-176(9)

Publisher: John Wiley & Sons, Ltd.

Abstract:

BACKGROUND: Samples from different plant species, different organs or tissues at different times of the year, usually show great differences in their cell compositions, pH, and the endogenous RNase activities, decreasing the RNA yield and quality.

RESULTS: In this study we describe a reagent and a simple total RNA isolation method for plant organs, tissues and dry seeds. The RNA extraction reagent (MAGi) is non-toxic and can be stored at room temperature for several months to years. The principle of the total RNA extraction is that tissues are lysed in extraction solution with the aid of mortar homogenization-maceration, and cellular proteins, polysaccharides and DNA are removed from the RNA. We tested the reported method on more than 16 different types of plant seed and 15 different tissues and organs of pepper.

CONCLUSION: The RNA extraction procedure reported in the present study greatly reduces the time required to isolate dry seed total RNA and other tissues by more than half as compared with the previously reported methods. The range of typical RNA yield and quality represents a significant improvement over existing protocols. The quality is high enough to be considered as suitable method for RT-PCR, cDNA library construction and microarray gene expression studies. Copyright © 2008 Society of Chemical Industry

Keywords: dry and immature seeds; RNA extraction reagent; MAGi; RT-PCR

Document Type: Research article

DOI: http://dx.doi.org/10.1002/jsfa.3428

Publication date: 2009-01-15

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