PCR primers (Table S2) were generated using Primer3 software (http://www-genome.wi.mit.edu/cgi-bin/primer/primer3_www.cgi). Amplicons were designed to display sizes of 80 bp–400 bp. Five microliters PCR reactions were performed using 3 ng–10 ng of genomic DNA, 2.5 mM MgCl2, 0.1 mM dNTP, 0.025 U AmpliTaq Gold DNA polymerase, and 1 pmol of eight mixed primers. Initial denaturation at 95° for 10 min was followed by 40 cycles of 95° for 30 s, 55° for 15 s, and 72° for 30 s using GeneAmp PCR Biosystem 9700 in 384-well reaction plates. PCR conditions were verified by direct DNA sequencing using mixed PCR products after treatment with exonuclease (USB), 0.5 units per sample, and shrimp alkaline phosphatase (SAP, USB), 0.50 U/sample for 30 min at 37°, and 5 min at 95°. Following multiplex DNA amplification, unincorporated PCR primers and dNTPs were degraded by adding 0.5 units per sample exonuclease and 0.50 U/sample SAP, incubating for 30 min at 37° and incubating for a further 5 min at 95°. After purification, 1.5 μl of cleaned PCR product was mixed with 5.5 μl of water, 2.0 μl
