Bijli, K.M. and Singh, B.P. and Sridhara, S. and Arora, N. (2001) Isolation of Total RNA from Pollens. Preparative Biochemistry and Biotechnology , 31 (2). pp. 155-162.
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Abstract
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 from pollens. To find a simple and reliable method for isolating total RNA from pollen, four methods, viz. phenol/SDS (PS), guanidine HCl (GH), tri-reagent (TR), and modified SDS-βME (SB) were tested with fresh pollen of Ricinus communis (procured at −70°C) and pollen dried at 30-37°C. The quality and quantity of RNA was superior for the material processed at −70°C. SB gave the highest RNA yield (2.35 mg/g, OD260/280 > 2.0), compared to other methods. The results obtained by the SB method were found to be comparable with the widely used tri-reagent method. This was validated with other pollens of Imperata cylindrica and Xanthium strumarium. The yield obtained from graded amounts of pollen was consistent with SB, compared to the TR method. The RNA isolated by SB gave good quality mRNA for synthesizing cDNA. The SDS-βME method is simple, efficient, and uses less expensive reagents. Hence, we recommend the modified SDS-βME method for isolating total RNA from pollens.
| Item Type: | Article |
|---|---|
| Author Affiliation: | Centre for Biochemical Technology, Delhi University Campus, Delhi |
| Subjects: | Crop Improvement |
| Divisions: | General |
| Depositing User: | Mr. SanatKumar Behera |
| Date Deposited: | 27 Jun 2012 11:05 |
| Last Modified: | 27 Jun 2012 11:05 |
| Official URL: | http://dx.doi.org/10.1081/PB-100103381 |
| URI: | http://eprints.icrisat.ac.in/id/eprint/6264 |
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