Modified Protocol for Isolation of High Quality RNA from the Matured Bark Tissue of tossa Jute

Abstract

Abstract Jute is classified as a bast fibre crop which is one of the most important natural fibre crops. Jute fibre is both economical and environmentally friendly and it can be utilized for minimizing the use of synthetic fibres. For the quality improvement of jute fibre, the molecular study of the genes linked with the fibre biogenesis is crucial. RNA isolation is a fundamental step for gene expression and transcriptomic studies of jute fibre. RNA isolation is restricted from jute especially from the well-developed bark tissues because it is rich in mucilage, polysaccharides, and phenolic compounds. In this study, a new method was optimized for extracting the total RNA from the field-grown bark tissues of two tossa jute varieties at 30 and 60 Days-After-Germination (DAG). Two-times use of CTAB extraction buffer with SDS and later addition of TRIZOL made the protocol simple, cost effective and minimal time consuming as compared to other methods. The A260/A280 value ranged between 1.99 ± 0.05 to 2.10 ± 0.02 which determined the purity of RNA. The range of the mean yield of the RNA was 232.38 ± 5.01 to 419.49 ± 9.43 µg/ gm of tissue which is abundant for further molecular analysis. For validation of the protocol, the RNA was converted into cDNA which was further amplified by semi quantitative-real-time-PCR using Cellulose synthase gene specific primers. The results indicated that the proposed RNA isolation method will be helpful in further downstream processing for the betterment of jute fibre quality.

Keywords

• Jute
• Fibre
• RNA isolation
• cDNA
• Ces