As. Pac. J. Mol. Biol. & Biotech., July 2012 Vol. 3, 107-113
Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis
Chai Har Siah1, Parameswari Namasivayam2,3 and Rozi Mohamed1*
1Forest Biotech Laboratory, Department of Forest Management, Faculty of Forestry, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
2Department of Molecular & Cell Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
3Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
* Author for correspondence: Rozi Mohamed
Department of Forest Management, Faculty of Forestry, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
Tel.: 60-17-260 5392 (Mobile Phone), 60-3-8946 7183 (Office), Fax: 60-3-8943 2514, Email : This email address is being protected from spambots. You need JavaScript enabled to view it..
Abstract.
Aquilaria malaccensis (karas) produces oleoresin in its wood (known as agarwood) when responding to wounding and microbial infection. Wood tissues are known to contain high levels of polysaccharides, polyphenolics and secondary metabolites, which make RNA extraction challenging. In this work, six different methods for extracting RNA from wood tissues of A. malaccensis were compared. RNA yield, purity, and integrity number, were used as parameters to evaluate the efficiency of each method. Conventional methods yielded RNA with good purity but the RNA integrity was poor. The commercial RNeasy Plant Mini kit protocol was modified by means of scaling-up the reaction and combining all aliquots in the same RNeasy spin column, and yielded the highest yield while maintaining the integrity of the RNA. We found that this kit with some modifications was most suitable for extracting RNA from healthy wood and agarwood. This study is essential for future molecular studies on agarwood.
