In the recent years, substantial progress has been made in the characterization of the rhesus macaque MHC region, which is more complex than in humans. To cope with the increasing knowledge, improved typing techniques for MHC genes are required. Including the DNA-sequences in this report, 39 rhesus macaque (Mamu)-DQB1 alleles, corresponding to 38 deduced protein sequences, are known. Here, we present a typing technique for Mamu-DQB1 alleles and the DNA-sequences of 9 novel DQB1 alleles. The technique consists of one or two rounds of screening followed by DNA-sequence determination. The first round represents a low-resolution screening sufficiently insensitive to identify novel alleles, which also allows sequencing of both alleles in most heterozygous individuals. The second round consists of a high-resolution screening. This is only necessary for animals in which one PCR-product was generated by the initial screening. The technique was validated by analyzing samples of more than 200 rhesus macaques of different origin, and DNA-sequence determination of more than 230 PCR-products.