Actually, I am trying for months to make a primer of a Clubroot resistance gene (CRa). CRa gene is found in three Brassica species. My aim is to make a primer that can amplify this one gene in all three species.
It is easy to make the primer of a gene. first, blast the gene with your desired species genes. desired gene sequence could use the primer3plus browser. choose the best optimum primer.
Hari Shankar Gadri , I made around 50 primers using these genes and sequece, only one primer worked. I am again trying that primer with same PCR conditions but there is no result.