But Primer3 is not a "point-and-click" black box. It is a . To unlock its full potential, you must understand its input format.
PRIMER_PRODUCT_SIZE_RANGE=150-250 PRIMER_PRODUCT_OPT_SIZE=200 primer3 input
Next time you are frustrated that "no primers were found," don't tweak the sequence—. Loosen the Tm range by 1°C or extend the product size range. The perfect primer pair is always just a parameter change away. Have a tricky multiplexing scenario? Drop the parameters in the comments below. But Primer3 is not a "point-and-click" black box