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[skyenemy]

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 away 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....

Edited by skyenemy, 17 December 2009 - 03:05 AM.