大麥 RFLP marker의 開發

Development of RFLP markers in barley(Hordeum vulgare L.)

Genomic single or low copy number DNA clones from random genomic DNA libraries using the plasmid vector pBR322 and the phage λgtll were constructed using DNA from a barley cultivar, Sacheon #6. This work was done to develop the barley RFLP DNA markers and to saturate the linkage groups. In the initial screening, 25 clones, which were expected to have single or low-copy sequence, were selected from 1,200 bacterial colonies and 50 genetically unique phage plaques. Of these, 10 clones showed polymorphisms. To do these works, DNAs that were isolated from the barley lines and cultivars, MMS, Apex, Olbori, Sacheon #6, were digested with 3 restriction enzymes, Bam HI, Hind Ⅲ and Eco RI to prepare the genomic blot. Chemiluminescence as well as nick-translation was utilized to label the probe.