Sequence-Tagged Site(STS) marker is a kind of important marker for identifying heterogeneous chromosome on account of its simplicity, speediness and specificity. There are many STS markers in use, which changed from the RFLP and RAPD markers. But some of those from RFLP markers cannot directly be used for identifying the specific chromosomes, such as MWG913. Amplified wheat genome by MWG913, a STS marker of barley's 1H chromosome, then cloned the products. These clones were classified by restricted enzyme Taq I and then sequenced. On the basis of the results of these sequences compared with barley's (obtained from the GenBank)., two restricted enzyme specifying on 1H chromosome, EcoR I and Pvu I, were chosen to digest the PCR products of T. aestivum, Secale cereale, Hordeum vulgare, Agropyron intermedium, Haynaldia villosa, Thinopyrum elongatum and identified their specilization to Hordeum vulgare genome. Furthermore, according to the difference of Pvu I recognition site, the second PCR primers were designed and identified its specificity to Hordeum vulgare genome by PCR. Thus 1H chromosomespecific CAPs and ASA markers of barley were obtained.