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Kumar G.R.K.,Plant Metabolic Engineering Group | Eswaran N.,Plant Metabolic Engineering Group | Johnson T.S.,Plant Metabolic Engineering Group
Analytical Biochemistry | Year: 2011

A method for isolating transcriptionally active RNA for downstream applications from diverse tissues of Jatropha curcas, a plant rich in latex, lipids, waxes, polysaccharide, polyphenols, and secondary metabolites, is described. The described method uses alkaline borate buffer during tissue homogenization to negate the formation of viscous gel observed in guanidium-salt-containing methods. By this method, quality RNA was extracted from leaf, immature inflorescence, endosperm, and root tissues with yields ranging from 1.80 to 7.80 mg/100 mg fresh weight (FW). The total RNA obtained was found to be suitable for poly(A)+RNA purification, complementary DNA (cDNA) synthesis, cloning of full-length cDNA, and cDNA library construction. © 2011 Elsevier Inc. All rights reserved.


PubMed | Plant Metabolic Engineering Group
Type: Journal Article | Journal: Analytical biochemistry | Year: 2011

A method for isolating transcriptionally active RNA for downstream applications from diverse tissues of Jatropha curcas, a plant rich in latex, lipids, waxes, polysaccharide, polyphenols, and secondary metabolites, is described. The described method uses alkaline borate buffer during tissue homogenization to negate the formation of viscous gel observed in guanidium-salt-containing methods. By this method, quality RNA was extracted from leaf, immature inflorescence, endosperm, and root tissues with yields ranging from 1.80 to 7.80mg/100mg fresh weight (FW). The total RNA obtained was found to be suitable for poly(A)(+)RNA purification, complementary DNA (cDNA) synthesis, cloning of full-length cDNA, and cDNA library construction.

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