Evaluation of a Flow-Through Depuration System to Eliminate the Human Pathogen Vibrio Vulnificus from Oysters
he reaction mixture for the first/single step PCR contained crude DNA extract (template DNA), reaction buffer (100mM Tris with 15mM MgCl2), 0.4Âµg each of forward and reverse primer (1s5 & 1a16; IK1 & IK2), 100mM each dNTPs (deoxyribonucleotide triphosphates), 3U Taq DNA polymerase and molecular grade water. The components were mixed thoroughly and the PCR reaction was conducted in the Thermalcycler (GeneAmp 9700, ABI Systems, Rotkreuz, Switzerland). The PCR protocol comprised of 35 cycles of 60 sec at 94Â°C, 60 sec at 55Â°C and 90 sec at 72Â°C. The programme included an initial delay of 4 min at 94Â°C and final extension of 5 min at 72Â°C before and after 35 cycles, respectively.