2.3. Allelic Frequency and Gene Diversity Analysis

Allele frequencies were estimated according to Bernstein's method (1925) [23] from the phenotypic data; the expected frequency was calculated under the assumption of the Hardy-Weinberg equilibrium from Rh and ABO phenotypes, with the Expected-Maximization (EM) algorithm [24]. Gene diversity was analyzed according to Nei [25]. The Nei genetic distances [26] were calculated based on the gene frequencies data of the ABO and D loci, and a dendrogram was constructed using the neighbor-joining (NJ) clustering procedure with the POPTREEW software [27]. The gene frequencies were used for the Principal Components Analysis (PCA).