Notice of RetractionEstablishment of an Ecotilling Protocol with M13 Primers for SNP Discovery in Barley

Notice of Retraction

After careful and considered review of the content of this paper by a duly constituted expert committee, this paper has been found to be in violation of IEEE´s Publication Principles.

We hereby retract the content of this paper. Reasonable effort should be made to remove all past references to this paper.

The presenting author of this paper has the option to appeal this decision by contacting TPII@ieee.org.

Ecotilling is the application of Tilling (Targeting Induced Local Lesions in Genomes) for discovery of nucleotide polymorphisms in natural populations, but this technique with M13 primers is not established in barley yet. The purpose of this study is to develop an Ecotilling protocol with M13 primers suitable to discover nucleotide polymorphisms for barley natural population. A set of 66 barley accessions originated from 19 countries were selected as natural population, the nested PCR was employed to amplify the target fragment of a chlorophyll a/b binding protein gene (Cab) by using gene specific primers and M13 primers labeled with IRD fluorescence dyes. An Ecotilling protocol which includes template amount and annealing temperature in PCR amplification, CEL I nuclease amount and digestion time in mismatch cleavage was optimized and established. Following the optimized protocol, six nucleotide polymorphisms were unambiguously discovered in the natural population, indicating the established Ecotilling protocol can effectively detect nucleotide polymorphisms with lower cost, and it could provide a foundation for larger scale efforts in characterization of natural nucleotide variation in barley.