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A simple and effective method for RNA isolation and cDNA library construction from the brown seaweed Sargassum polycystum (Fucales, Phaeophyceae)

The isolation of high-quality RNA from brown seaweeds has always been problematic due to a high content of polyphenolic compounds and polysaccharides in their tissues. This study presents a simple and effective method for isolating high-quality RNA from the brown seaweeds Sargassum (Sargassum polycy...

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Main Authors: Sim, Mei Chea, Ho, Chai Ling, Phang, Siew Moi
Format: Article
Language:English
Published: Springer 2013
Online Access:http://psasir.upm.edu.my/id/eprint/28106/1/A%20simple%20and%20effective%20method%20for%20RNA%20isolation%20and%20cDNA%20library%20construction%20from%20the%20brown%20seaweed%20Sargassum%20polycystum%20%28Fucales%2C%20Phaeophyceae%29.pdf
http://psasir.upm.edu.my/id/eprint/28106/
http://link.springer.com/article/10.1007%2Fs10811-013-9980-z
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spelling my.upm.eprints.281062016-06-20T06:39:24Z http://psasir.upm.edu.my/id/eprint/28106/ A simple and effective method for RNA isolation and cDNA library construction from the brown seaweed Sargassum polycystum (Fucales, Phaeophyceae) Sim, Mei Chea Ho, Chai Ling Phang, Siew Moi The isolation of high-quality RNA from brown seaweeds has always been problematic due to a high content of polyphenolic compounds and polysaccharides in their tissues. This study presents a simple and effective method for isolating high-quality RNA from the brown seaweeds Sargassum (Sargassum polycystum, Sargassum siliquosum, Sargassum baccularia, and Sargassum binderi), Padina tetrastromatica, Turbinaria conoides, Dictyota sp., and Hormophysa cuneiformis using cetyltrimethylammonium bromide and selective lithium chloride precipitation. Approximately 25–43 μg g−1 fresh weight of total RNA was obtained from these brown algae. The A 260/A 280 absorbance ratio of these RNA samples are between 1.995 ± 0.027 and 2.122 ± 0.013, with distinct 28S and 18S ribosomal RNA bands visible on a formaldehyde agarose gel. The RNA obtained from S. polycystum was found to be suitable for many downstream applications such as reverse transcription polymerase chain reaction, cloning, synthesis of cDNA, and construction of cDNA library for the generation of expressed sequence tags. Springer 2013 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/28106/1/A%20simple%20and%20effective%20method%20for%20RNA%20isolation%20and%20cDNA%20library%20construction%20from%20the%20brown%20seaweed%20Sargassum%20polycystum%20%28Fucales%2C%20Phaeophyceae%29.pdf Sim, Mei Chea and Ho, Chai Ling and Phang, Siew Moi (2013) A simple and effective method for RNA isolation and cDNA library construction from the brown seaweed Sargassum polycystum (Fucales, Phaeophyceae). Journal of Applied Phycology, 25 (5). pp. 1277-1285. ISSN 0921-8971; ESSN: 1573-5176 http://link.springer.com/article/10.1007%2Fs10811-013-9980-z 10.1007/s10811-013-9980-z
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description The isolation of high-quality RNA from brown seaweeds has always been problematic due to a high content of polyphenolic compounds and polysaccharides in their tissues. This study presents a simple and effective method for isolating high-quality RNA from the brown seaweeds Sargassum (Sargassum polycystum, Sargassum siliquosum, Sargassum baccularia, and Sargassum binderi), Padina tetrastromatica, Turbinaria conoides, Dictyota sp., and Hormophysa cuneiformis using cetyltrimethylammonium bromide and selective lithium chloride precipitation. Approximately 25–43 μg g−1 fresh weight of total RNA was obtained from these brown algae. The A 260/A 280 absorbance ratio of these RNA samples are between 1.995 ± 0.027 and 2.122 ± 0.013, with distinct 28S and 18S ribosomal RNA bands visible on a formaldehyde agarose gel. The RNA obtained from S. polycystum was found to be suitable for many downstream applications such as reverse transcription polymerase chain reaction, cloning, synthesis of cDNA, and construction of cDNA library for the generation of expressed sequence tags.
format Article
author Sim, Mei Chea
Ho, Chai Ling
Phang, Siew Moi
spellingShingle Sim, Mei Chea
Ho, Chai Ling
Phang, Siew Moi
A simple and effective method for RNA isolation and cDNA library construction from the brown seaweed Sargassum polycystum (Fucales, Phaeophyceae)
author_facet Sim, Mei Chea
Ho, Chai Ling
Phang, Siew Moi
author_sort Sim, Mei Chea
title A simple and effective method for RNA isolation and cDNA library construction from the brown seaweed Sargassum polycystum (Fucales, Phaeophyceae)
title_short A simple and effective method for RNA isolation and cDNA library construction from the brown seaweed Sargassum polycystum (Fucales, Phaeophyceae)
title_full A simple and effective method for RNA isolation and cDNA library construction from the brown seaweed Sargassum polycystum (Fucales, Phaeophyceae)
title_fullStr A simple and effective method for RNA isolation and cDNA library construction from the brown seaweed Sargassum polycystum (Fucales, Phaeophyceae)
title_full_unstemmed A simple and effective method for RNA isolation and cDNA library construction from the brown seaweed Sargassum polycystum (Fucales, Phaeophyceae)
title_sort simple and effective method for rna isolation and cdna library construction from the brown seaweed sargassum polycystum (fucales, phaeophyceae)
publisher Springer
publishDate 2013
url http://psasir.upm.edu.my/id/eprint/28106/1/A%20simple%20and%20effective%20method%20for%20RNA%20isolation%20and%20cDNA%20library%20construction%20from%20the%20brown%20seaweed%20Sargassum%20polycystum%20%28Fucales%2C%20Phaeophyceae%29.pdf
http://psasir.upm.edu.my/id/eprint/28106/
http://link.springer.com/article/10.1007%2Fs10811-013-9980-z
_version_ 1643829371353432064
score 13.211869

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