SNP-RFLP - problem with artifical restriction site (Dec/17/2009 )

for SNP-RFLP, i am making an artifical resistriction site on pcr product by changing primer sequence, because no enzyme is available in original sequence.

introducing 2 mismatches(5 nucleotides awau from the SNP site) into the primer is better,
or just introducing 1 mismatch in the primer (but the mismatch is just 2 nucleotide away from SNP site)

someone told me that the mismatch in primer should not be closed to the 3'end of primer...
but i dunt have choice, only a few restriction enzymes can cut the SNP........


anyone can give me opinion??
i am just a undergraduate(Bsc.), and i am suffered by the project....

Hi..
Maybe This attachment can help for you.I didn't more research about this issue.