Optimization of DNA Isolation and RAPD Technique in Arecanut (Areca catechu L.)
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Date
2007
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Abstract
A simple and efficient protocol for extracting high quality DNA from arecanut (Areca catechu L.) leaves is presented. DNA yield and purity were monitored by gel electrophoresis and by determining absorbance at UV (A260/A280). The ratio was between 1.7 to 1.9 indicating that the presence of contaminating metabolites was minimal. The quantities of DNA obtained were 100- 400 ig/g starting material. The isolated DNA proved amenable to PCR amplification and restriction digestion. DNA was completely digested with the three restriction enzymes (EcoR I, EcoR V and Hind III) confirming the purity of the extracted DNA. Using the isolated DNA, the parameters for randomly amplified polymorphic DNA (RAPD) protocol was standardized.
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Keywords
Arecanut palm, gel electrophoresis, enzimes, EcoR I, EcoR V, Hind III
Citation
Agrotrópica 19: 31 - 34. 2007.