Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica

To survive in extremely cold environments, psychrophilic microorganisms produce exopolysaccharides (EPS), which are carbohydrate polymers that constitute a substantial component of the extracellular polymers surrounding cells of these microorganisms. EPS can interfere with RNA extraction and decre...

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Main Authors:	Izwan, B., Najad-Zamirah, Z., Farah-Diba, A.B., Nor-Muhammad, M., Mohd-Nazalan, M.N, Rosli, M.I., Abdul-Munir, A.M.
Format:	Article
Language:	English
Published:	Penerbit Universiti Kebangsaan Malaysia 2014
Online Access:	http://journalarticle.ukm.my/8681/ http://journalarticle.ukm.my/8681/ http://journalarticle.ukm.my/8681/1/43_2_09.pdf

id	ukm-8681
recordtype	eprints
spelling	ukm-86812016-12-14T06:47:52Z http://journalarticle.ukm.my/8681/ Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica Izwan, B. Najad-Zamirah, Z. Farah-Diba, A.B. Nor-Muhammad, M. Mohd-Nazalan, M.N Rosli, M.I. Abdul-Munir, A.M. To survive in extremely cold environments, psychrophilic microorganisms produce exopolysaccharides (EPS), which are carbohydrate polymers that constitute a substantial component of the extracellular polymers surrounding cells of these microorganisms. EPS can interfere with RNA extraction and decrease the purity of the RNA extracted from EPS producing microorganisms. In this work, six commercial RNA extraction kits and two published protocols for RNA extraction were evaluated for total RNA extraction from the psychrophilic yeast, Glaciozyma antarctica. All the protocols were optimised to obtain the highest quality of total RNA. The results show that all of the tested commercial kits and the tested conventional methods yielded RNA from G. antarctica, albeit with varying quality. The protocol that utilises TRIzol® reagent was the most effective method for isolating total RNA from G. antarctica of which this protocol resulted in the highest RNA yield and purity compared to other methods. This method of RNA extraction produced RNA of sufficient quality for reverse transcriptase PCR (RT-PCR) to detect the expression of the G. antarctica delta 9-fatty acid desaturase gene as well as for the construction of a G. antarctica cDNA library. Penerbit Universiti Kebangsaan Malaysia 2014-12 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/8681/1/43_2_09.pdf Izwan, B. and Najad-Zamirah, Z. and Farah-Diba, A.B. and Nor-Muhammad, M. and Mohd-Nazalan, M.N and Rosli, M.I. and Abdul-Munir, A.M. (2014) Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica. Malaysian Applied Biology, 43 (2). pp. 71-79. ISSN 0126-8643 http://mabjournal.com/index.php?option=com_content&view=article&id=460&catid=59:current-view&Itemid=56
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Kebangasaan Malaysia
building	UKM Institutional Repository
collection	Online Access
language	English
description	To survive in extremely cold environments, psychrophilic microorganisms produce exopolysaccharides (EPS), which are carbohydrate polymers that constitute a substantial component of the extracellular polymers surrounding cells of these microorganisms. EPS can interfere with RNA extraction and decrease the purity of the RNA extracted from EPS producing microorganisms. In this work, six commercial RNA extraction kits and two published protocols for RNA extraction were evaluated for total RNA extraction from the psychrophilic yeast, Glaciozyma antarctica. All the protocols were optimised to obtain the highest quality of total RNA. The results show that all of the tested commercial kits and the tested conventional methods yielded RNA from G. antarctica, albeit with varying quality. The protocol that utilises TRIzol® reagent was the most effective method for isolating total RNA from G. antarctica of which this protocol resulted in the highest RNA yield and purity compared to other methods. This method of RNA extraction produced RNA of sufficient quality for reverse transcriptase PCR (RT-PCR) to detect the expression of the G. antarctica delta 9-fatty acid desaturase gene as well as for the construction of a G. antarctica cDNA library.
format	Article
author	Izwan, B. Najad-Zamirah, Z. Farah-Diba, A.B. Nor-Muhammad, M. Mohd-Nazalan, M.N Rosli, M.I. Abdul-Munir, A.M.
spellingShingle	Izwan, B. Najad-Zamirah, Z. Farah-Diba, A.B. Nor-Muhammad, M. Mohd-Nazalan, M.N Rosli, M.I. Abdul-Munir, A.M. Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica
author_facet	Izwan, B. Najad-Zamirah, Z. Farah-Diba, A.B. Nor-Muhammad, M. Mohd-Nazalan, M.N Rosli, M.I. Abdul-Munir, A.M.
author_sort	Izwan, B.
title	Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica
title_short	Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica
title_full	Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica
title_fullStr	Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica
title_full_unstemmed	Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica
title_sort	comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, glaciozyma antarctica
publisher	Penerbit Universiti Kebangsaan Malaysia
publishDate	2014
url	http://journalarticle.ukm.my/8681/ http://journalarticle.ukm.my/8681/ http://journalarticle.ukm.my/8681/1/43_2_09.pdf
first_indexed	2023-09-18T19:52:57Z
last_indexed	2023-09-18T19:52:57Z
_version_	1777406340928372736

Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica

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