To develop the STS (Sequence Tagged Site) marker for discrimination between oat cultivars used the EEG (Euchromatin Enriched Genomic) DNA library in oat. The EEG DNA library was constructed the Mcr A and Mcr BC system in DH5 alpha bacterial cell line. About 3,500 EEG colonies had been constructed by using junk DNA exclusion. About 800 colonies were selected that included insert DNA more than 500 bp. It was analyzed the genetic information by using blast searches of NCBI web site. More than three hundred STS primer sets were developed using sequencing data of selected colonies and about 90 primer sets which showed single band were selected in Olgwiri. It was applied to twelve oat cultivars including Olgwiri and has been shown polymorphism at 15%. PCR product which amplified with selected STS primer was treated with six endonucleases and was showed polymorphic bands. These primers could be useful for specific allele tagging in mapping populations and germplasm and for the study of functional genomics of oat.
The purpose of this study is to develop the EEG (Euchromatin Enriched Genomic) DNA library of wheat, barley, rye and oat. Mcr A and Mcr BC system in DH5 alpha bacteria cell line and Kuemkangmil, Olbori, Olhomil and Olgwiri were used for materials in our experiments. EEG colonies have been constructed by using junk DNA exclusion. We analyzed the genetic information of the colonies using blast searches of NCBI and GRAMENE web sites. One hundred eighty-four, 65, 79 and 119 STS primer pairs were developed using sequencing data of selected colonies in Kuemkangmil, Olbori, Olhomil and Olgwiri respectively. Twenty-eight and forty-two percent of designed primer pair showed polymorphism using six endoucleases in Kuemkangmil, Olbori, Olhomil and Olgwiri germplasm respectively. These primers could be useful for specific allele tagging in mapping populations and germplasm and for the study of functional genomics of wheat, barley, rye and oat.