Amplification reactions were performed on a GeneAmp® PCR System 9700 (9700 cycler) (AB) or a 24-PIKO® Thermal Cycler (PIKO cycler) (Bioké). In the PIKO cycler appropriate ultra-thin walled tubes were used (Bioké). SGM+, Identifiler and SEfiler+ amplifications used 28 cycles, while NGM amplifications took 29 cycles.
Standard reactions used amplification conditions as provided by the manufacturer. Phusion amplifications on the 9700 cycler consisted of 1 min 98 °C hotstart, 28 cycles of 5 s 98 °C denaturation, 20 s 59 °C primer annealing and 20 s 72 °C elongation with a final soak of 30 min 60 °C. Unless stated otherwise, rapid amplification reactions on the PIKO cycler employed an initial denaturation at 98 °C for 5 min, followed by the appropriate number of cycles at 98 °C for at least 5 s, 59 °C for at least 30 s, 72 °C for 10 s with a final extension at 72 °C for 1 min.