Dna isolation

Horsák, Michal & Horsáková, Veronika, 2025, Unveiling Icelandic land snail fauna: new discoveries and biogeographical insights, Contributions to Zoology 94 (3), pp. 292-306 : 294-295

DNA isolation , alignment and genetic analyses

The identification of the species and the determination of their probable biogeographical origin were approached by ana- lysing their mitochondrial DNA . DNA was isolated from the whole specimens using the E.Z.N.A. Mollusc DNA Kit (Omega Bio-tek). PCR amplification was performed using the primers listed in table 1. PCR products were purified using ExoSAP (Affymetrix) and sequenced at SEQmes.r.o. (CZ). The laboratory protocols, sequence assembly and alignment preparation fol- low our routine protocols published in Horsáková et al. (2019). Protein-coding genes were translated into amino acids to check for stop codons. The resulting align- ments were partitioned by gene and, in the case of protein-coding genes, by first + second and third codon position.For each par- tition, optimal substitution models were

CONTRIBUTIONS TO ZOOLOGY 94 (2025) 292–306

identified in jModelTest v. 2.1.10 ( Darriba et al., 2012), and Bayesian inference (BI) was performed in MrBayes v3.2.6 ( Huelsenbeck & Ronquist, 2001) with the settings described in detail in Horsáková et al. (2019). To visualise the relationships between the Icelandic mtDNA haplotypes and other haplotypes recorded in Europe, haplotype networks were created using the TCS algorithm ( Clement et al., 2000) in PopART v.1.7 ( Leigh & Bryant, 2015). Available sequences from populations in the distribution ranges of each target species and their phylogenetically closely related taxa were retrieved from GenBank. The GenBank accession numbers of the newly obtained sequences are listed in table 2.