Alpha-Gliadin amplicon of Bread Wheat
Amplicon was sequenced by the Illumina MiSeq system producing 2 x 300 paired-end reads. Illumina amplicon library was prepared and sequenced at the Unidad de Genmica Cantoblanco of Fundacin Parque Cientfico de Madrid (FPCM, Spain) as previously reported (SnchezLen et al., 2018) with some modification. Briefly, to avoid bias of the sequencing results, a dilution of all samples considering the Cq were made to start the sequencing with the same amount of -gliadin amplicon molecules, and  first PCR were terminated at exponential phase to maintain the proportionality of amplicon variants.  The range of amplicon lengths was check using the Agilent 2100 Bioanalyzer system (Agilent Technologies, Santa Clara, CA 95051).