based on the definition of tm, it follows that since binding will not (or at least should not) occur between the template and your added restriction site, you should not therefore factor it in when calculating tms, gc etc. Fyi, adding flag/myc tags etc on a primer will give you crazy tms. Again, no binding so dont worry about them. Gl
-- I sent this from my 3 mobile --
-original message-
Subject: tm calculation for tagged primers
From: "Azam Rahimpour" <
rahimpour_a@...>
Date: 02/11/2009 7:53 pm
Hi
I am new to primer design and I have a simple question: when I add extra sequences like restriction sites and epitope tags to my primer, should I calculate primer tm with these sequences or not? actually when I calculate Tm with these sequences the tm is about 80 and if I ignore them it seems that the calculation isnt complete.
is there any rule for that?
regards
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