2.4 Protocol 2: Assorted information of specimens with or without voucher exuviae
Culturing and morphological data. This method is clearly distinguished from Protocol 1 by retaining notably fewer data layers than Protocol 1. For example, while the WS-specimens are represented by all data layers, except for live images of the sequenced specimens, the AZ-material is exclusively represented by exuvial vouchers photographed in LM. Although admittedly artificial, we decided to lump all cases with varying degrees of morphological information, but no live imaging, into one category. Protocol 2 was applied to the three live-imaged specimens of H. demodex from Green Island (TA) in Olesen et al., (2022), the specimens of H. itoi from the White Sea (WS; for which all data layers except live-images of single specimens exist), and the Azores specimens (AZ; for which only y-naupliar exuviae are available).
2.4.1 DNA extraction. We tested three extraction procedures: that used exclusively for Protocol 1, the DNeasy™ kit (as described in detail above and in the Supplementary Material) following the manufacturer’s specifications, and the GeneReleaser™ kit (as described in detail above).