OPTIMIZATION OF GENOMIC DNA EXTRACTION FROM THE LEAF TISSUES OF JATROPHA SPECIES FOR MOLECULAR STUDIESV.D. MENDHULKAR AND HENA HAYAT
Depending on the nature and complexity of plant material, proper method needs to be employed for extraction of genomic DNA, along with its performance and evaluation by different molecular techniques. Jatropha a genus of family Euphorbiacea, known for its biofuel and medicinal properties. Jatropha species characteristically contains high concentrations of polyphenols, and polysaccharides that interfere with DNA, causing erroneous or no PCR products. In this study, genomic DNA was isolated from leaves of three species of Jatropha, namely J. curcas, J. podagrica and J. gossipyfolia. We optimized and employed a simple genomic DNA isolation protocol suitable for Jatropha plants from leaves. The DNA obtained from leaf tissues with a modified cetyltrimethylammonium bromide buffer and using no liquid nitrogen protocol was of good quality, with no colored pigments and contaminants. The spectral quality of genomic DNA isolated using this method as measured by the A260/A280 absorbance ratio ranged from 1.80 to 1.91 and the concentration were found to be 8.60, 9.45 and 9.50 (μg/μL) for the three speciess of Jatropha. The amplified plants DNA extracted from leaves of Jantropha plants using modified CTAB buffer method with no liquid nitrogen were successfully amplified by PCR using arbitrary RAPD primers.
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