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To partly address these shortcomings, we used the method developed by Bretman & Tregenza [ 36] which estimate the maximum likelihood number of mating partners from population allele frequencies.
The difference in the maximum likelihood number of mating partners between female morphs was analysed using a Mann Whitney U test.
The frequency of positive and negative reactions was used to Poisson estimate the maximum likelihood number of amplifiable molecules and its 95% confidence intervals [Jeffreys et al., 1994].
Both estimates of polyandry (number of alleles, and maximum likelihood number of mating partners), were consistent with a smaller number of mating partners in androchrome females [ 36] Figure 2, although the estimate for maximum likelihood number of mating partners per morph type should be interpreted with caution since it is based on a small sample size of 3 individuals.
To estimate the maximum likelihood number of mating partners of females from Louro (N = 17), the DNA from the muscle thorax of 15 males was extracted and genotyped for the six microsatellites, following the protocol explained in previous section.
Based on male allele frequencies in Louro, the maximum likelihood number of mating partners, which was estimated among 1 10 mating partners, for both morphs combined was 3.38 ± 1.82 SD (N females from Louro = 17, range = 1 8).
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Assuming estimated rates of digit gain, maximum likelihood numbers of pedal digits for all nodes are consistent with those for the Bayesian majority-rule consensus (see Additional file 1) in more than 90% of trees.
Maximum likelihood numbers of manual and pedal digits for internal nodes were determined independently using Mesquite [ 26], assuming the asymmetrical Markov k-state 2-parameter (AsymmMk) model of character evolution.
For all nodes, maximum likelihood numbers of manual digits reconstructed for more than 90% of trees are consistent with (i.e., identical to or only insignificantly more likely than) those inferred assuming the Bayesian majority-rule consensus (see Figure 1), and reversals of limb reduction are inferred for only an insignificant proportion (less than 5%) of trees.
When using more than one gene, the maximum-likelihood number of partners for each female is estimated using the locus showing the highest degree of putative partners.
We refer to the copy number best supported by the combined data as the "Maximum Likelihood Copy Number" (MLCN), and the factor by which the support for that copy number exceeds the next best copy number (the "minimum ratio", MR) is an index of confidence in the assignment of the copy number.
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