Centre : Central Potato Research Institute(CPRI)
Name of the Protocol
Simultaneous detection of Fusarium spp and Phytophthora infestans
Nature : Novel
Crop / Pathogen : Potato/ Fusarium spp and Phytophthora infestans.
PCR reaction was performed in a total volume of 25 µl, contacting of 1X GreenTaq Buffer (with 1.5mM MgCl2), 200µM of each dNTPs, both pathogen specific primers 10µM(F+R), Taq DNA Polymerase Enzyme(5U/ µl) ), 20ng/µl Template DNA and remaining milliQ water. The PCR reaction was performed in thermal cycler (Eppendorff Mastercycler).The PCR conditions are as follows: denaturation at 94 °C for 2 min, followed by 30 cycles of 94 °C for 30 second, 55 °C for 30 second, 72 °C for 1 min and a final extension at 72 °C for 10 min. Amplified PCR products (600 bp for Phytophthora infestans & 700 bp for Fusarium spp) were separated by electrophoresis on 2.0 % agarose gel and picture was captured under gel documentation system.