OPTIMIZATION OF DNA ISOLATION PROTOCOL IN SELECTED CULTIVARS OF SESAMUM INDICUM L.
T.N. DHAR *
Cytogenetics and Plant Biotechnology Research Unit, Department of Botany, University of kalyani, Kalyani, Nadia-741235
C. SEN GUPTA *
Cytogenetics and Plant Biotechnology Research Unit, Department of Botany, University of kalyani, Kalyani, Nadia-741235
*Author to whom correspondence should be addressed.
Abstract
Genetic analysis of plant is relished on high yield of pure DNA sample. In this paper we present the optimization of DNA isolation protocol of selective cultivar of Sesamum indicum. The method involves a modified CTAB extraction employing PVP yield grinding. Successive long term chloroform: isoamyl alcohol extraction and RNAase treatment with all steps carried out at room temperature. The yield of DNA ranges from 205.2 to 721.2 μg/ml., 231.6 to 781.3 μg/ml and 218.1 to 768.8 μg/ml. of leaf tissue and the purity was 1.65 to 1.79, 1.68 to 1.85 and 1.60 to 1.87 indicating minimal level of contaminating particles. The techniques are ideal for DNA isolation and can be used for further molecular studies.
Keywords: Sesamum indicum, CTAB, Benzyl Chloride, PVP, Agarose gel, DNA isolation.