Genomic DNA was extracted from whole blood from rhesus macaques (Macaca mulatta) from the NIH Animal Center (NIHAC), and direct sequencing was performed using samples from 20 unrelated animals (pairwise Identity by Descent, or IBD ≤ 0.0125) that were selected on the basis of variable HPA axis activity. We used primers designed from published human sequence and, subsequently, from rhesus sequence generated in our lab (available upon request). Cycle sequencing was performed using the Big Dye Terminator Version 3.1 reaction in 96-well optical plates (Applied Biosystems, Inc., Foster City, CA). Variants were detected by visualization of electropherograms generated by ABI Sequencing Analysis software.
