PCR amplification and pyrosequencing

High resolution KIR genotyping of DNA samples from Iraqw, Datooga, Baka, Mbuti and Hadza individuals was performed as described (23). Specific amplicons corresponding to individual KIR gene exons were made and subjected to pyrosequencing using a PSQ HS 96A instrument (Qiagen). Novel KIR alleles found in the African populations, were validated by generating an independent PCR amplicon, cloning each allele and sequencing several clones by Sanger methodology.