The fit between observed (O) and expected (E) frequencies is statistically contrasted by a χ2 test, since the summation Σ(O − E)2/E is distributed as a χ2 distribution with a number of degrees of freedom equal to the number of genotypic classes that are compared less the number of parameters that are needed to obtain the expected values. In this case there are six classes and to obtain the expected values two of the three allele frequencies are needed (the third is determined once the first two are known) and the total number of individuals in the sample. Therefore, the number of degrees of freedom is d.f. = 6 − 2 − 1 = 3.
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