Fig. 2

Filtration functions of Ultiplex. Primers need to be filtered with different functions to eliminate malfunctional pairs. A Getprimers.harpin_filter function. If there is a hairpin structure in the combined sequence of the primer and tag, the primer pair will be eliminated. B Getprimers.dimer_filter function. If there is a dimer structure between any two combined primer and tag sequences, the primer pair will be eliminated. C Getprimers.area_filter & Getprimers.site_filter functions. If any skipped sites or areas are located at the 3′ end of the primer, the primer pair will be eliminated. D Getprimers.single_blast_filter function. The potential binding sites of a single primer are evaluated, and if the amplicon length is below the cut-off value, the primer pair will be eliminated