SNPs and indels were selected for validation by traditional Sanger sequencing. Primers were designed using Primer3 (http://frodo.wi.mit.ed/primer3/). PCR assays were performed using 5 ng DBA/2J genomic DNA, 10 pmol each of forward and reverse primer in 50 μl. The following cycle parameters were used: 95 °C for 4 min; 35 cycles of 95 °C for 30 s; 55 °C for 30 s and 72 °C for 1 min; and 72 °C for 5 min. PCR products were purified with 2 μl ExoSAP-IT (Invitrogen Corporation). Sanger sequencing was performed using an ABI 3730.
