The likelihood is , and a maximum likelihood estimator is any maximizer . Write and score . Fisher information for the sample is
where the equality uses the usual differentiability and interchange assumptions. For independent identically distributed observations . Under standard interior, identifiability and regularity assumptions,
Without those assumptions the large-sample normal assertion need not hold, for example at a boundary parameter.
The totals are independent binomials, and . Thus
Differentiating the log likelihood, or maximizing the two factors separately, gives
This remains the constrained maximum at zero or one when a total is at an endpoint. At interior parameters the sample information, in coordinate order , is
For example ; the cross derivative is zero. As both sample sizes grow with nondegenerate interior probabilities, the joint approximation is normal with mean and covariance . The independence of the two estimators is exact, inherited from the two data samples.
A linear combination of independent normal random variables is normal, with variances adding when the coefficients are . Applying the joint central limit approximation from (i) gives
Equivalently the difference minus , divided by the displayed standard deviation, converges in distribution to a standard normal under the same regular large-sample regime.
Under the null put . The likelihood becomes the same two binomial constants times . Maximization gives the pooled estimate
Under an interior null, (ii) has zero mean and variance . Statistical consistency of and Slutsky's theorem permit replacement of by in the denominator, giving the stated . For a two-sided level- test, reject when . The large-sample approximation requires both expected counts of each type to grow; if all observed alleles are identical the stated denominator degenerates and this approximation is not applicable.
At level the two-sided critical values are . Since , do not reject the two-sided null. The approximate two-sided -value is .
For the specified alternative , the statistic should be negative. Reject in the lower tail, . Since , reject for this one-sided alternative, with approximate -value . The direction of the tail follows the order of subtraction in the statistic.

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