sample PCRs, were checked on 2% agarose gels (SeaKem LE, Cambrex, ME, USA). For cycle sequencing, we used Big Dye Terminator Mix v3.1 (Applied Biosystems, Foster City, CA) at 0.33 µl of mix per reaction in a total volume of 4 µl with 50 cycles of amplification and ABI 3730 capillary sequencers. DNA sequence for 14 out of 24 genes was analyzed with Phred/Phrap/polyphred-5.02/polyphred-7/Consed 14 as described [47]. The remainder of genes was analyzed using Mutation Surveyor (Softgenetics, PA, USA). 5 genes were analyzed with both software tools to demonstrate no appreciable difference in the detection of sequence variants. In total we generated and analyzed 34.8 million base pairs. All variants were verified by at least two researchers.
