title: Isolation of RNA from field-grown jute (Corchorus capsularis) plant in different developmental stages for effective downstream molecular analysis.
abstract: Jute (Corchorus capsularis), as a natural fibre producing plant species, ranks next to cotton only. Today, biotechnological approach has been considered as most accepted means for any genetic improvement of plant species. However, genetic control of the fibre development in jute has not yet been explored sufficiently for desired genetic improvement. One of the major impediments in exploring the genetic architecture in this crop at molecular level is the availability of good quality RNA from field-grown plant tissues mostly due to the presence of high amount of mucilage and phenolics. Development of a suitable RNA isolation method is becoming essential for deciphering developmental stage-specific gene expression pattern related to fibre formation in this crop species. A combination of modified hot borate buffer followed by isopycnic centrifugation (termed as HBIC) was adopted and found to be the best isolation method yielding sufficient quantity (~350-500 μg/gm fresh tissue) and good quality (A(260/280) ratio 1.88 to 1.91) RNA depending on the developmental stage of stem tissue from field-grown jute plant. The poly A(+) RNA purified from total RNA isolated by the present method was found amenable to efficient RT-PCR and cDNA library construction. The present development of RNA isolation was found to be appropriate for gene expression analysis related to fibre formation in this economically important jute plant in near future.
authors.affiliation: Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology, Kharagpur 721302, India.
authors.forename: Pradipta
authors.initials: P
authors.lastname: Samanta
authors.forename: Sanjoy
authors.initials: S
authors.lastname: Sadhukhan
authors.forename: Subrata
authors.initials: S
authors.lastname: Das
authors.forename: Alpana
authors.initials: A
authors.lastname: Joshi
authors.forename: Soumitra K
authors.initials: SK
authors.lastname: Sen
authors.forename: Asitava
authors.initials: A
authors.lastname: Basu
mesh_terms: D032381:Corchorus / Q000235:genetics / Q000254:growth & development*; D004587:Electrophoresis, Agar Gel; D015723:Gene Library; D018547:Plant Stems / Q000737:chemistry* / Q000254:growth & development; D018749:RNA, Plant / Q000032:analysis / Q000302:isolation & purification*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013784:Textiles
chemical_list: D018749:RNA, Plant