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> The Tarpy papers recently cited suggested 12 +/- 6, but there is no mention of how this is determined. Does anyone know?
> Examining sperm is the norm.
The method of determining mating number in 2021 is:
> Once the queens had mated and started to lay eggs (roughly 4–6 weeks following the introduction of their cells), we collected ~ 200 emerging worker offspring from each surviving nucleus colony and stored them in separate vials of 70% ethanol. We used the worker samples for genotyping analysis to measure mating frequency of each queen.
> Genotyping analysis. We molecularly analyzed each worker by removing one hind leg, placing it in a 0.2 mL strip tube, and cutting it up with scissors that were ethanol and flame-sterilized between samples. DNA was extracted from each sample with either the Chelex 100 (BioRad) or HotSHOT protocols.
> We detected relatively high polyandry overall, ranging from 25 to 38 subfamilies per colony. Our current estimates of mating number (mean ± SD: 30.9 ± 4.41) are on par with or even exceed the average mating frequencies for Apis mellifera.
Rusert, L.M., Pettis, J.S. & Tarpy, D.R. Introduction of Varroa destructor has not altered honey bee queen mating success in the Hawaiian archipelago. Sci Rep 11, 1366 (2021). https://doi.org/10.1038/s41598-020-80525-5
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