Harm-4, found in Uganda and Australia, was the most diverged haplotype with seven mutation steps from the major haplotype Harm-1 (Fig. 1).
The network analysis revealed three distinct sets of haplotypes which were separated by more than 23 mutation steps from each other.
The other haplotypes were located within the IRS2 and were separated by at least 30 mutation steps from each other and by 24-26 mutations from the closest IRS1 haplotype.
Interestingly, there was no any single nonsynonymous mutation detected in, at least, 10 mutation steps from the Zp3cdh1 to the two outgroup species, i.e., the domestic sheep and goat.
Our ancient sequences were also similar to this common extant haplotype with a single mutation step away from it, thus suggesting genetic continuity between Norway spruce populations.
The addition of the four STRs DYS437, DYS438, DYS459a, and DYS455 showed no further variation while the addition of DYS459b and DYS454 demonstrated two additional haplotypes that were each comprised of two samples one mutation step away from the extended CMH.
To test the significance of this grouping, we performed AMOVA analysis by classifying 217 Indian animals into two groups, viz.; 1) twenty-five animals with one mutation step away from the node R1, and b) all the remaining Indian buffalo as a separate group.
Moreover, 30 haplotypes were definable with a single mutation difference from H1, with H2 seven mutation steps removed from H1 (Fig. 3).
The mutation step size is drawn from a normal distribution of mean zero and standard deviation σ μ.
For instance the first amino acid in the triresidue from early chordates is alanine (encoded by GCG), which undergoes two synonymous mutations leading to the corresponding actual residue in mammalian H2A.Z-2 (alanine, GCT), with an intermediate synonymous mutation step represented by H2A.Z-2 from amphibians (Xenopus, alanine GCC).
