title: Improved and convenient method of RNA isolation from polyphenols and polysaccharide rich plant tissues.
abstract: It has been difficult to extract a good quality total RNA from the plant parts (such as seeds) which contain high levels of phenolic compounds, carbohydrates and other compounds that bind and/or co-precipitate with RNA. A simple, rapid and efficient method for isolating total RNA from polyphenols and polysaccharide rich plant tissues has been developed. Seeds of leguminosae family were chosen for the study. The good quality and high yield of total RNA was achieved with A260/A280 ratio of 1.9. Seeds of three different crops (Cajanus cajan, Dolichos biflorus and Vigna mungo) at different developmental stages were evaluated for total RNA extraction using standardized protocol. Seeds at 21 days after flowering (DAF) gave the best results among others (7 DAF and dry seeds). Quality of isolated RNA from all the three crops was further checked by cDNA synthesis. The extracted RNA was found suitable for further molecular applications such as reverse transcription and cDNA library construction.
authors.affiliation: National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi 110 012, India. rekhakansal@hotmail.com
authors.forename: Rekha
authors.initials: R
authors.lastname: Kansal
authors.forename: Kalika
authors.initials: K
authors.lastname: Kuhar
authors.forename: Isha
authors.initials: I
authors.lastname: Verma
authors.forename: Ram Niwas
authors.initials: RN
authors.lastname: Gupta
authors.forename: Vijay Kumar
authors.initials: VK
authors.lastname: Gupta
authors.forename: Kirpa Ram
authors.initials: KR
authors.lastname: Koundal
mesh_terms: D005591:Chemical Fractionation / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics; D007887:Fabaceae / Q000737:chemistry* / Q000235:genetics*; D005419:Flavonoids / Q000032:analysis*; D010636:Phenols / Q000032:analysis*; D059808:Polyphenols; D011134:Polysaccharides / Q000032:analysis*; D018749:RNA, Plant / Q000302:isolation & purification*
chemical_list: D018076:DNA, Complementary; D005419:Flavonoids; D010636:Phenols; D059808:Polyphenols; D011134:Polysaccharides; D018749:RNA, Plant