A primer binding site mutation at the D2S1338 locus resulting in a loss of amplification

Abstract

An apparent paternal genetic incompatibility at the D2S1338 locus due to false homozygosity is described. It was disclosed after amplification with the AmpFlSTR ® Identifiler ® PCR Amplification Kit and reproducible with the genRES ®MPX-4 DNA-Typing Kit. After application of an alternative primer set an allele *25 could be amplified, which restored Mendelian inheritance between father and child. Sequencing revealed a C > T transition at the fourth position from the 3′ end of the standard forward primer, which potentially caused the allelic loss with the primer sets in the commercial multiplex PCR kits.