title: An improved protocol for the isolation of RNA from roots of tea (Camellia sinensis (L.) O. Kuntze).
abstract: Tea, a beverage crop, is a rich source of polyphenols and polysaccharides which greatly attribute to its importance. However, oxidation and precipitation of these compounds during nucleic acids extraction is a limitation to molecular biology and genomic studies. On isolation of total RNA from root tissue using established protocols, difficulties were encountered in terms of purity and quantity of isolated RNA or some of the methods were time-consuming and also yields were low. The present communication combines a phenol-based RNA isolation protocol with a cetyltrimethylammonium bromide-based procedure with appropriate modifications. This protocol successfully isolated RNA from tap root tissue in 2-3 h as compared with 16 h reported by the previous method. Also, RNA yield was higher by more than fourfold. The RNA isolated by this protocol was successfully used for downstream applications such as RT-PCR and the construction of suppression subtractive hybridization library. The developed protocol worked well with other plant tissue with high polyphenols and polysaccharides contents.
authors.affiliation: Biotechnology Division, Council of Scientific and Industrial Research-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh 176061, India.
authors.forename: Richard Chalo
authors.initials: RC
authors.lastname: Muoki
authors.forename: Asosii
authors.initials: A
authors.lastname: Paul
authors.forename: Anita
authors.initials: A
authors.lastname: Kumari
authors.forename: Kashmir
authors.initials: K
authors.lastname: Singh
authors.forename: Sanjay
authors.initials: S
authors.lastname: Kumar
mesh_terms: D028241:Camellia sinensis / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D008967:Molecular Biology / Q000379:methods*; D018517:Plant Roots / Q000235:genetics*; D018749:RNA, Plant / Q000302:isolation & purification*; D013662:Tea / Q000737:chemistry*
chemical_list: D018749:RNA, Plant; D013662:Tea