ddRADseq data from 94 segregating individuals and parents generated from an F7 isofemale family from a cross between FANG and FUMOZ colones

Female progeny phenotyped according to the bioassay test from two isofemale families at the F2 and F7 generation originating from a FUMOZ female crossed with a FANG male were selected for ddRADseq. For each family, 96 samples were sequenced, including 94 females (48 resistant and 46 susceptible) and parental FANG and FUMOZ females. ddRADseq libraries were produced using an IGATech (Udine, Italy) custom protocol, with minor modifications to Peterson's double digest restriction-site associated DNA preparation. Genomic DNA was fluorometrically quantified, normalized to a uniform concentration and 75ng are double digested with 2.4U of both NlaIII and MluCI endonucleases (New England BioLabs) in 30µL reaction supplemented with CutSmart Buffer and incubated at 37°C for 90', then at 65°C for 20'. Fragmented DNA was purified with 1.5 volumes of AMPureXP beads (Agencourt) and subsequently ligated with 200U of T4 DNA ligase (New England BioLabs) to 2pmol of both overhang barcoded adapters for NlaIII and MluCI cut sites in 40µL reaction incubated at 23°C for 60' and at 20°C for 60' followed by 20' at 65°C. Samples were pooled into multiplexed batches and bead purified as before. For each pool, targeted fragments distribution is collected on BluePippin instrument (Sage Science Inc.) setting the range of 400 - 520 bp using a 2% agarose cassette. Gel eluted fractions were amplified with indexed primers using Phusion High-Fidelity PCR Master Mix (New England BioLabs) in as final volume of 50µL and subjected to the following thermal protocol [95°C, 3'] - [95°C, 30'' - 60°C, 30'' - 72°C, 45''] x 12 cycles - [72°C, 2']. Products were then purified with 1 volume of AMPureXP beads. The resulting libraries were checked with both Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA) and Bioanalyzer DNA assay (Agilent Technologies, Santa Clara, CA). Libraries were then sequenced with 150 cycles in paired-end mode on NovaSeq 6000 instrument following the manufacturer's instructions (Illumina, San Diego, CA).