水稻中随机扩增多态性DNA（RAPD）标记的测定及其向序列标签位点（STS）和STS特异性引物的转化。
Determination of RAPD markers in rice and their conversion into sequence tagged sites (STSs) and STS-specific primers.
作者信息
Monna L, Miyao A, Inoue T, Fukuoka S, Yamazaki M, Zhong H S, Sasaki T, Minobe Y
机构信息
Rice Genome Research Program, National Institute of Agrobiological Resources, Ibaraki, Japan.
出版信息
DNA Res. 1994;1(3):139-48. doi: 10.1093/dnares/1.3.139.
We produced 102 randomly amplified polymorphic DNA (RAPD) markers mapped on all 12 chromosomes of rice using DNAs of cultivars Nipponbare (japonica) and Kasalath (indica) and of F2 population generated by a single cross of these parents. Sixty random primers 10 nucleotides long were used both singly and in random pairs and about 1,400 primer-pairs were tested. Using both agarose gel and polyacrylamide gel electrophoresis enabled us to detect polymorphisms appearing in the range from < 100 bp to 2 kb. The loci of the RAPD markers were determined onto the framework of our RFLP linkage map and some of these markers were mapped to regions with few markers. Out of the 102 RAPD markers, 20 STSs (sequence-tagged sites) and STS-specific primer pairs were determined by cloning, identifying and sequencing of the mapped polymorphic fragments.
我们利用日本晴（粳稻）和卡萨拉斯（籼稻）品种的DNA以及由这两个亲本单交产生的F2群体，在水稻的所有12条染色体上产生了102个随机扩增多态性DNA（RAPD）标记。使用了60个10个核苷酸长的随机引物，单独使用和随机配对使用，共测试了约1400对引物。同时使用琼脂糖凝胶电泳和聚丙烯酰胺凝胶电泳，使我们能够检测出大小从小于100bp到2kb范围内出现的多态性。将RAPD标记的位点确定在我们的RFLP连锁图谱框架上，其中一些标记被定位到标记较少的区域。在102个RAPD标记中，通过对定位的多态性片段进行克隆、鉴定和测序，确定了20个序列标签位点（STS）和STS特异性引物对。
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