Phusion® Flash polymerase (Phusion) (Bioké, Leiden, the Netherlands) was tested for direct PCR. A typical reaction had a 20 μL reaction volume and consisted of 10 μL 2× enzyme-containing Master Mix, 4 μL of appropriate STR primer mix and 6 μL of water and DNA. Standard AmpliTaq Gold® DNA Polymerase (AmpliTaq) (AB) amplification reactions were carried out in 25 μL reaction volumes. Set-up occurred as recommended by the manufacturer.
The following autosomal STR kits were used: SGM+, AmpFℓSTR® Identifiler® (Identifiler), AmpFℓSTR® SEfiler Plus™ (SEfiler + ) or AmpFℓSTR® NGM™ (NGM) (all AB).