Comments

Warning: This is a very basic tool and should not be seen as a replacement for your brain. This is in no way a complete tool for primer design and shouldn't be trusted! Always check the results yourself. The Tm calculation is based on Belsauer et al. 1986 (with some small modifications, inspired by http://www.basic.northwestern.edu/biotools/oligocalc.html) and has as a goal to stay close (in principle it should be the same) to the results found by the NEB Phusion HF-PCR reference (the kit we're using). The tail is ignored in the Tm calculation. Also take a look at iGEM's primer design page.

Known limitations are: only A, C, G and T are accepted as input, the concentration of salts(50mM of Na+), and the primer concentration (500nM) can't be set.