Phenotypes and genotypes of brown trout used for breeding experiments in 2014

Standardized DNA extracts were sent to Ecogenics GmbH (Balgach, Switzerland) for microsatellite typing at 13 loci (i.e. Brun13, Brun25, BS131, MST-15, Mst543, MST-591, Ssa-197, Ssa-85, SSOSL438, Str12INRA, Str2INRA, Str58CNRS and T3-13) using in house protocols (see Supplementary Material of Palejowski et al. 2022 for full details). Amplification of microsatellite loci was conducted in three multiplex reactions. Multiplex one and two, each containing primers for the amplification of four microsatellites, was performed using 2X QIAGEN® Multi-plex PCR Master Mix (Qiagen GmbH, Hilden, Germany). Multiplex three included primers for the amplification of five microsatellite loci and was amplified using 2X HotStarTaq Mastermix (Qiagen GmbH, Hilden, Germany). Before thermal cycling, a prolonged denaturation step (95°C for 15 min) was included followed by 35 cycles with 94°C for 30 s, 58°C for 90 s, 72°C for 60 s and a final elongation step of 30 min at 72°C. Fragment analyses were performed on a 3730XL DNA Analyzer (Applied Biosystems, Foster City, CA, USA) with a GeneScan LIZ500 size standard (Applied Biosystems) and allele calling was performed using the GeneMarker V2.6.4 software (SoftGenetics LLC, State College, PA, USA). Palejowski H., Bylemans J., Ammann V., Marques da Cunha L., Nusbaumer D., Castro I., Uppal A., Mobley K.B., Knörr S., Wedekind C. 2022 Sex-specific life history affected by stocking in juvenile brown trout. Front. Ecol. Evol. 10, 869925. (doi:10.3389/fevo.2022.869925).