i am designing primer for SNP-RFLP and no restriction enzyme is available to cut the SNP site.
so, i make an artifical restriction site by inducing a primer mismatch in the primer, so that the pcr product can be cut by enzyme.
however, the mismatch i induced is near the SNP site, and after cutting by enzyme, the fragment is too short that cannot be visualized...
someone said i can add a poly(T) tail to the 5' end of the mismatch primer so that the fragment can be long enough.
anyone know whether it will work???
primer design problem
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