title: Isolation of poly (A)+ mRNA for downstream reactions from some medicinal and aromatic plants.
abstract: In the present protocol for extraction of RNA, hexadecyltrimethylammoniumbromide (CTAB) and insoluble polyvinylpyrrolidone were used followed by LiCl precipitation, CsCl ultracentrifugation and finally poly (A)+ mRNA was isolated with the help of oligo(dT)-cellulose columns. The isolated poly (A)+ mRNA was found to be suitable for cDNA-AFLP and suppression subtractive hybridization applications. It is a modified and consolidated protocol based on previously described methods for isolated steps and works better for medicinal and aromatic plants. High yield of poly (A)+ mRNA coupled with its amenability for downstream reactions like RT-PCR, northern blotting and cDNA synthesis for library construction is a key feature of the present protocol.
authors.affiliation: Genetic Resources and Biotechnology Division, Central Institute of Medicinal and Aromatic Plants, Lucknow 226 015, India.
authors.forename: Ashutosh K
authors.initials: AK
authors.lastname: Shukla
authors.forename: Ajit K
authors.initials: AK
authors.lastname: Shasany
authors.forename: Suman P S
authors.initials: SP
authors.lastname: Khanuja
mesh_terms: D030008:Catharanthus / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry*; D012333:RNA, Messenger / Q000302:isolation & purification*; D018749:RNA, Plant / Q000302:isolation & purification*; D012015:Reference Standards
chemical_list: D010936:Plant Extracts; D012333:RNA, Messenger; D018749:RNA, Plant