OPTIMASI AMPLIFIKASI PCR (Polymerase Chain Reaction) SEKUEN GEN matK cpDNA PADA TANAMAN MANGGA (Mangifera) RIAU

Abstract

The ability of mango from Sumatra to bloom in high rainfall season makes this plant potential to be superior germplasm. Conservation of superior germplasm resources is necessary to produce superior mango species. The study of molecular taxonomy has an important role in conservation and management of plant genetic resources. Molecular taxonomy studies are supported by PCR technique as a basic technique in molecular research. This study aimed to obtain the optimal conditions to amplify the matK cpDNA gene of mango from Riau in order to obtained good PCR product. DNA isolation was conducted based on the modified CTAB method. The isolated DNA was then amplified using PCR technique at temperature of 95oC 4 min (pre-PCR), 95oC 30 seconds (denaturation), 49,1oC 30 seconds (annealing), 72°C 1,5 minutes (extension), 72°C 10 minutes (post-PCR) as many 35 cycles. PCR optimization has been done by varying the components of the PCR, such as the variations volume of DNA template (0.5 μl, 1 μl, 2 μl and 4 μl) and Taq PCR (Hot Start Green and DreamTaq Green). Application of DreamTaq Green and 1 μl template DNA showed the best amplification results of gene matK cpDNA which was marked with thick and no smear band of DNA