Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis

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 di...

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Main Authors: Siah, Chai Har, Namasivayam, Parameswari, Mohamed, Rozi
Format: Article
Language:English
Published: Malaysian Society for Molecular Biology and Biotechnology 2012
Online Access:http://psasir.upm.edu.my/id/eprint/58500/
http://psasir.upm.edu.my/id/eprint/58500/1/Comparisons%20of%20different%20RNA%20extraction%20methods%20on%20woody%20tissues%20of%20the%20tropical%20tree%2C%20Aquilaria%20malaccensis.pdf
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author Siah, Chai Har
Namasivayam, Parameswari
Mohamed, Rozi
author_facet Siah, Chai Har
Namasivayam, Parameswari
Mohamed, Rozi
author_sort Siah, Chai Har
building UPM Institutional Repository
collection Online Access
description 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.
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spelling upm-585002018-01-19T07:54:06Z http://psasir.upm.edu.my/id/eprint/58500/ Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis Siah, Chai Har Namasivayam, Parameswari Mohamed, Rozi 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. Malaysian Society for Molecular Biology and Biotechnology 2012 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/58500/1/Comparisons%20of%20different%20RNA%20extraction%20methods%20on%20woody%20tissues%20of%20the%20tropical%20tree%2C%20Aquilaria%20malaccensis.pdf Siah, Chai Har and Namasivayam, Parameswari and Mohamed, Rozi (2012) Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis. Asia Pacific Journal of Molecular Biology and Biotechnology, 20 (3). pp. 107-113. ISSN 0128-7451 http://www.msmbb.org.my/apjmbb/html203/203e.htm
spellingShingle Siah, Chai Har
Namasivayam, Parameswari
Mohamed, Rozi
Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis
title Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis
title_full Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis
title_fullStr Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis
title_full_unstemmed Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis
title_short Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilaria malaccensis
title_sort comparisons of different rna extraction methods on woody tissues of the tropical tree, aquilaria malaccensis
url http://psasir.upm.edu.my/id/eprint/58500/
http://psasir.upm.edu.my/id/eprint/58500/
http://psasir.upm.edu.my/id/eprint/58500/1/Comparisons%20of%20different%20RNA%20extraction%20methods%20on%20woody%20tissues%20of%20the%20tropical%20tree%2C%20Aquilaria%20malaccensis.pdf