Nice program, looks like it might save me some time. In
regards to real time primers have you tested that the
primers generated work? If so what machine did you use,
what method and how efficient are they?
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I've designed about 30 pair of real time primers and all but
one pair works fine with a 3 step program on a Cepheid
smartcycler. Efficiencies range from 82%-108% with most in
I also have extremely good experiences using perlprimer
(using a 7500 instrument). I designed ~20 primer pairs and
all have worked.
I think that it would be really great if you implemented a feature where you could specify the min. intron size for intron spanning primers.