DOI: 10.5061/dryad.7h44j0zwj
title: Phenotypes and genotypes of brown trout used for breeding experiments in 2014

description: Adult brown trout were caught via electrofishing around the beginning of
 the spawning season from the River Aare and its tributaries Gürbe, Worble,
 Giesse, Kiese, and Rotache (Bern canton, Switzerland). They were kept in
 the Fischereistützpunkt Reutigen until eggs could be stripped from
 females. Adults were narcoticised when ready to spawn and gametes were
 stripped for block-wise full-factorial in vitro fertilisations (e.g.
 Marques da Cunha et al. 2019). Photographs were taken from all fish,
 standard length and wet weight were determined, and fin clips were taken
 for molecular analyses. All fish were then released back into the streams
 of origin. Samples of 24 freshly fertilized eggs per experimentally
 produced full-sib family were used for various laboratory experiments
 (e.g. Marques da Cunha et al. 2019). The remaining embryos were raised in
 the Fischereistützpunkt Reutigen and used for stocking the streams of
 parental origin and various nursery streamlets, following the routine of
 the stocking program of the Bern canton. These stocked fish were later
 sampled at various stages of their life cycle, and 13 microsatellite
 markers could be used to assign these fish to their parents. Marques da
 Cunha L., Uppal A., Seddon E., Nusbaumer D., Vermeirssen E.L.M., Wedekind
 C. 2019. No additive genetic variance for tolerance to ethynylestradiol
 exposure in natural populations of brown trout (Salmo trutta). Evol. Appl.
 12(5), 940-950. (doi:10.1111/eva.12767).
description: 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. <em>Front. Ecol. Evol</em>. 10,
 869925. (doi:10.3389/fevo.2022.869925).  
description: These fish were used to
 experimentally produce embryos in a large number of full-factorial
 breeding blocks. Subsamples of these embryos were used in various types of
 laboratory experiments or released into different streamlets and sampled
 at various stages.