croplm

Study Extent

Specimens were collected at eight sampling sites in Medvednica Nature Park.

Sampling

Adults were collected using an entomological net while larvae were collected by hand under stones and branches.

Quality Control

All collected individuals were preserved in ethanol. Morphological determination of adults was conducted using a ZEISS SteREO Discovery V20 stereo microscope along with various identification keys. Due to the lack of identification keys the larvae were not determined to the species level. Larval identification was carried out by comparing their sequences with sequences available in databases.

Method steps

1. Total genomic DNA was isolated from the tissues of one to three legs of adults and larvae using the GenElute Mammalian Genomic DNA Miniprep Kit. Chromatograms were analyzed and edited in Geneious Prime. Misread nucleotides were manually corrected. The presence of stop codons in all sequences was checked using Mesquite. Poor quality sequences and those containing stop codons were excluded from further analyses. For phylogeographic analyses, we selected species with a broader distribution and a larger number of COI sequences available in the BOLD and GenBank databases.

Stoneflies

1. Plecoptera
   common name: Stoneflies rank: order

Croatia (Medvednica Mt.)