The β-globin gene cluster has shown high polymorphic diversity organized in 5′ and 3′ haplotypes (Hps). β S-Chromosomes are in linkage disequilibrium with the 5′ Hps Bantu, Benin, Senegal, Cameroon, and Arab–Indian. In Mexican mestizos with African west coast origins, we observed the following 5′ Hps in β S-chromosomes: Bantu, 78.8%; Benin, 18.2%; and atypical Hp 9, 3.0%. With the purpose of establishing the 3′ Hps, we analyzed 35 polymorphic sites—6 by RFLP analysis and 29 by DNA sequencing—in 33 unrelated β S-chromosomes. The polymorphic sites were structured according to Harding et al. [R.M. Harding, S.M. Fullerton, R.C. Griffiths, J.B. Clegg, Archaic African and Asian lineages in the genetic ancestry of modern humans, Am. J. Hum. Genet. 60 (1997) 772–789] and Lapouméroulie et al. [C. Lapouméroulie, O. Dunda, R. Ducrocq, G. Trabuchet, M. Mony-Lobé, J.M. Bodo, P. Carnevale, D. Labie, J. Elion, R. Krishnamoorthy, A novel sickle cell mutation of yet another origin in Africa: the Cameroon type, Hum. Genet. 89 (1992) 333–337]. All Bantu β S-chromosomes showed the 12A1 3′ Hp with (AT) 6T 9 repeats (84.9%), a novel 3′ Hp. The Benin Hp was 2B2, with (AT) 8T 4 (12.1%), and the atypical Hp 9 4B1, (AT) 8T 5 (3.0%). Because of the high linkage disequilibrium observed for the Bantu and 12A1 Hps, we expect that, if there is a single origin of the Bantu β S mutation, all must show the 12A1 polymorphic DNA sequence in the 3′ Hp. A correlation between the 5′ and 3′ Hps could be observed with the other β S mutations. The atypical Hp 9 was also atypical at the 3′ Hp, with the same repeats as observed with the Cameroon β S mutation; however, it differed in one position from the typical Lapouméroulie Cameroon Hp, indicating that these β S-chromosomes arose by different genetic mechanisms or by a novel β S mutation. We stress the importance of the study of DNA polymorphisms at 3′ Hp to allow understanding of the genetic diversity of β S-chromosomes, as well as their implications in β S gene expression and the possible effects on the clinical phenotype.