Use of universal reporter primers in multiplex PCR of autosomal loci

Abstract At present forensic identification and parentage testing is invariably carried out using multiplex PCR amplification kits in which many loci, whether they be STRs, SNPs or DIPs are analyzed simultaneously using primers labelled with at least three, but usually four, different fluorophores. Should existing loci need to be substituted by newer, for instance, more polymorphic loci, new fluorescent primers need to be synthesized; indeed, the dye labels of several loci may need to be switched to allow a re-accommodation of the allele sizes in the single multiplex, and this can be costly. We sought to facilitate these substitutions and reduce costs by using four universal primers each labelled with a different fluorophore and employ these in a test multiplex PCR amplification of 20 DIP loci.