Primer3 is a widely used program for designing PCR primers (PCR = "Polymerase Chain Reaction"). PCR is an essential and ubiquitous tool in genetics and molecular biology. Primer3 can design hybridization probes and sequencing primers. PCR is used for many different goals. Consequently, primer3 has many different input parameters that you control and that tell primer3 exactly what characteristics make good primers for your goals. Primer3 picks primers for PCR reactions, considering as criteria: * oligonucleotide melting temperature, size, GC content and primer-dimer possibilities, * PCR product size * positional constraints within the source (template) sequence * possibilities for ectopic priming (amplifying the wrong sequence) * many other constraints. All of these criteria are user-specifiable as constraints, and some are specifiable as terms in an objective function that characterizes an optimal primer pair. manual online: https://primer3.org/manual.html For details and citation see "References" in the package's doc folder.