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Next-generation sequencing of amplicons is a rapid and reliable method for the detection of polymorphisms relevant for barley breeding

Fritsch, Leonie, Soeur, Raphael, Hansen, Claudia, Fischer, Rainer, Schillberg, Stefan, Schröper, Florian
Molecular breeding 2016 v.36 no.7 pp. 83
Hordeum vulgare, barley, crops, genotype, genotyping, high-throughput nucleotide sequencing, plant breeding, polymerase chain reaction
Modern plant breeding is based on the detection of traits at the molecular level so that the selection process can be accelerated. Methods that allow the rapid and high-throughput analysis of such traits are therefore useful for plant breeders and farmers. We have shown that next-generation sequencing, applied to short PCR products spanning or flanking polymorphisms of interest in barley, can provide robust genotyping data that allow the rapid determination of genotype and zygosity. This method can be used to genotype large panels of plants because up to 80 million individual reads can be produced in one sequencing run, and samples from different lines and/or traits can be pooled. These findings are significant because plant breeders may need to screen large populations for multiple traits in parallel. Our next-generation sequencing method therefore provides a simple and inexpensive approach for the rapid and accurate genotyping of natural polymorphisms in barley, which can also be applied in many other economically relevant crop species.