Isolation of total RNA from pollens

Isolation of total RNA from plant materials has been difficult, due to the presence of complex organic substances and the associated pigmentation. In fact, there is a dearth of standardized protocols for isolating total RNA f rom pollens. To find a simple and reliable method for isolating total RNA f rom pollen, four methods, viz. phenol/SDS (PS), guanidine HCl (GH), tri-rea gent (TR), and modified SDS-beta ME (SB) were tested with fresh pollen of R icinus communis (procured at -70 degreesC) and pollen dried at 30-37 degree sC. The quality and quantity of RNA was superior for the material processed at -70 degreesC. SE gave the highest RNA yield (2.35 mg/g, OD260/280 >2.0), co mpared to other methods. The results obtained by the SE method were found t o be comparable with the widely used tri-reagent method. This was validated with other pollens of Imperata cylindrica and Xanthium strumarium. The yie ld obtained from graded amounts of pollen was consistent with SE, compared to the TR method. The RNA isolated by SE gave good quality mRNA for synthesizing cDNA. The SD S-beta ME method is simple, efficient, and uses less expensive reagents. He nce, we recommend the modified SDS-beta ME method for isolating total RNA f rom pollens.