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Optimization of RAPD-PCR Protocol to Screen Jatropha Curcas and Gossypium Hirsutum Grown in Metal Contaminated Soil

Narayanan Mathiyazhagan and Devarajan Natarajan

The optimization of randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) protocol was performed to screen two plants ( Jatropha curcas & Gossypium hirsutum ) grown in metal contaminated soils. A CTAB method was employed for DNA extraction, but an overnight RNase treatment was carried out to eliminate RNA contaminants. The isolated DNA was used for RAPD-PCR amplification. The optimum condition for reliable amplification requires a higher concentration of MgCl 2 (3 mM), primer (2.5 μM), Taq DNA polymerase (1 unit) and 3μl of template DNA (sample) and an annealing temperature of 55°C. Reproducible amplified products were observed in these conditions for both the plant species.