Procedure 31 Rapid electrochemical verification of PCR amplification of Salmonella spp. based on m-GEC electrodes

Publisher Summary This chapter presents a procedure to perform a polymerase chain reaction (PCR) amplification of the IS200 insertion sequence of Salmonella spp. with double labeling of the amplicon by using labeled primer and quantify the double-labeling amplification products of Salmonella spp. by using an electrochemical strategy based on magnetic beads and m- graphite–epoxy composite (GEC) electrode. Amplification of the salmonella genome include DNA extraction, PCR reaction.DNA is extracted with phenol–chloroform–isoamyl alcohol (25:24:1) and DNA is by precipitated with isopropanol. The PCR is performed according to the kit manufacturer, in a 100 μL of reaction mixture containing PCR template, 200 μmol L –1 of each deoxynucleotide triphosphate, 0.5 μmol L –1 of each labeled primer and 5 U of Taq polymerase, and in buffer containing 1.5 μmol L–1 MgCl 2 .