Kinorhyncha COX1 sequences from Disko Island, Greenland

Material for the present study was collected from twelve sampling stations in August 3-9, 2023, during an expedition to Disko Island (Western Greenland), organised by the Natural History Museum of Denmark (NHMD). Sediment samples were collected with a dredge and/or a Van Veen grab. Kinorhynchs were extracted from the sediment, sorted under a stereomicroscope, preserved in 99 % EtOH and kept in −20 ◦C until further analysis. Single individuals were selected for DNA extraction using Tissue DNA Purification Kit (EURx, Gdansk, Poland) with additional step of cuticle retention (centrifugation of lysis buffer at 11,000×g, followed by separation of exoskeleton from supernatant including eluted DNA). The exoskeletons were recovered, transferred to a microscopic glass slide and mounted in Fluoromount-G for light microscopy (LM). The exoskeletons were examined using a Nikon 600 and an Olympus BX51 equipped with an Olympus SC300 camera, catalogued and deposited in NHMD. DNA samples were used for fragment cox1 gene PCR amplification using LCO1490 and HCO2198, forward and reverse primer pairs with final concentration 0.2 μM, respectively (Folmer et al., 1994). Amplification reaction was carried out using onHybrid PCR Master Mix (EURx, Gdansk, Poland) with the protocol consisting of: preliminary denaturation (5 min, 95 ◦C), 35 cycles (denaturation - 30 s, 95 ◦C; annealing - 30 s, 49 ◦C; extension - 90 s, 72 ◦C) and final extension (5 min, 72 ◦C). Samples were purified with PCR/DNA Clean-Up Purification Kit (EURx, Gdansk, Poland) and sent for EZ-Seq sequencing (Macrogen Europe). Obtained sequences were assembled into contigs, trimmed and analyzed using SeaView version 5.0.5 software (Gouy et al., 2010) and deposited in NCBI GenBank database with their assigned accession numbers.