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Optimisation of RNA extraction from Gracilaria changii (Gracilariales, Rhodophyta)
Authors:Cheong-Xin Chan  Swee-Sen Teo  Chai-Ling Ho  Rofina Yasmin Othman  Siew-Moi Phang
Institution:(1) Institute of Postgraduate Studies, University of Malaya, 50603 Kuala Lumpur, Malaysia;(2) Department of Biotechnology, Faculty of Food Science and Biotechnology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
Abstract:RNA extraction from seaweed tissues is problematic due to the presence of polysaccharides and polyphenolic compounds upon cell disruption. Besides, a successful RNA isolation from seaweed tissues can sometimes be strain- and species-specific. Four different methods were used to extract RNA from Gracilaria changii (Gracilariales, Rhodophyta), collected from the mangrove area at Morib, Selangor, Malaysia. An optimised and modified total RNA extraction method was developed for this recalcitrant species. The use of sand in tissue grinding, and the incorporation of phenol extraction at the initial stage resulted in the highest RNA yield (0.65–1.14 mgrg g–1 fresh weight) with high quality (A260:280 ratio 1.80–2.05). The RNA obtained is suitable for cDNA synthesis and future functional genomic studies.
Keywords:Gracilaria changii  Malaysia  polysaccharides  RNA extraction  seaweed
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