An improved method for RNA extraction from woody legume species Acacia koa A. Gray and Leucaena leucocephala (Lam.) de Wit

Author

Kazue Ishihara1, Eric K.W. Lee2, Dulal Borthakur3*

1,2,3*Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, 1955 East-West Road, Honolulu, HI 96822, United States.

Accepted December 12, 2015.

Citation: Ishihara K, Lee EKW, Borthakur D (2016). An improved method for RNA extraction from woody legume species Acacia koa A. Gray and Leucaena leucocephala (Lam.) de Wit. Int. J. Forestry Wood Sci. 3(1): 031-035.

Copyright: © 2016 Ishihara et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are cited.

Abstract

It is difficult to extract high-quality RNA of sufficient quantity from the stem tissues of tree legumes, such as Acacia koa and Leucaena leucocephala, because they contain high amounts of phenolic compounds and polysaccharides. The objective of this study was to develop an improved protocol that produces high-quality RNA from the stem tissues of these tree legumes. We developed a modified method that utilizes a lysis buffer containing a mixture of two commercially available reagents, RLT buffer from Qiagen RNeasy Kit and Fruit-mate™ from Takara. Comparison of the modified method with four other RNA extraction methods (Qiagen RNeasy, Fruit-mate™, TRIzol, and cetyltrimethylammonium bromide (CTAB)) showed that the modified method was far superior to the other methods. The extracted RNA produced reproducible DNA products in reverse transcription-polymerase chain reaction (RT-PCR). This improved protocol is rapid and easy, and it will facilitate genetic studies of A. koa, L. leucocephala, and other woody plants.

For further details log on website :
https://premierpublishers.org/ijfws/040220164808