restriction site in primers urgent - (Oct/17/2006 )

i have designed primers by the software of primer premier 5, now i want to add the restriction enzyme sites ,but i do not know how to do? guidlines for desinging reverse primer with restriction site (stop codon)? thanks in advance

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u have ur reverse primer and u also added the stop codon. before the stop codon, just copy the restriction seqeunce u wish to use (sites r palindromic). and u should have ur primer with Restriction site. Now add a few more bases (i add 5-6 A's) before the site so that u could digest it efficiently.



so I would have a reverse primer like this, for reverse primer

extra bases - xhoI site - stop codon - priming sequence
AAAAAA- CTCGAG - TTA -

hope this helps.

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