What are the recommendations for IDAA tri-primer design?

We recommend to use Primer3Plus (https://primer3plus.com/cgi-bin/dev/primer3plus.cgi), with primer lengths of 20-25 bp and G/C content of 30-70%. Primers should always be experimentally tested for specificity and efficacy using the standard tri-primer amplification parameters as descibed in Lonowski et al..