Multiplex PCR Genotyping of 46 AIMs

All individuals were genotyped for 46 AIMs [46 (link)] using a multiplex PCR amplification, as done in previous studies [25 (link),26 (link)]. In brief, each PCR reaction was a mixture of 5μl 2× Qiagen Multiplex PCR Master Mix, 1μl 10× Primer Mix, 0.5μl DNA (concentration between 0.5-5ng/μl), and 3.5μl water. The samples were prepared for capillary electrophoresis by adding 11.5μl Hi-Di Formamide (Applied Biosystems) and 0.3μl Liz-500 Size Standard (Applied Biosystems) to 0.8μl PCR product. A 3130xl Genetic Analyzer (Applied Biosystems) was used to separate DNA fragments by size. Analysis of indels was conducted by applying the software GeneMapper (Applied Biosystems). The genotyping results are reported in S5 Table.