jjw@rsbs0.anu.oz@ (10/23/90)
Hello Netlanders, I've just spent a little time designing some oligo primers to use in some PCR reactions for generating fragments with specific restriction sites on the ends. I searched high and low for information on what the minimum number of bases around a recognition site different R. enzymmes need for correct and efficient cutting. I found a reference on primer design for this purpose in a book on PCR which glibbly advises one to include a few extra bases 5'- to the restriction site being introduced but give no real info on how many are needed for which enzymees. So this I did and sent the sequences off to the university facillity for construction. Then I sat back and thought how nice it would be to have had the info. I needed; hence, this note. So much for the intro. I remember seeing *years ago!* some chart or table which had this sort of info. neatly listed. ie. - how many flanking nucleotides around a restriction site different R. enzymes need. Of course, I now don't remember where I saw this snippet of information. Does anybody out there remember where this table came out? Or where this info might be obtained? I think that the table I remember seeing was designed for the purpose of ensuring multiple enzyme digestions were compatible (ie. sites not too close together), but in this day and age this type of info. has great value in PCR primer design. So, ten brownie points for those with great memories who reply! Any many thanks in advance to those who don't remember the table I so dimly recall but who can suggest a likely souce of information. Cheers! ******************************************************************************** Jeremy Weinman Plant Microbe Interaction group Research School of Biological Sciences Australian National University Email: jjw@rsbs0.anu.oz Phone: 61 6 2495051 Fax: 61 6 2490754 Snail: PO Box 475, Canberra, ACT 2601, AUSTRALIA ********************************************************************************