Cushwa W T, Dodds K G, Crawford A M, Medrano J F
Department of Animal Science, University of California, Davis, California 95616-8521, USA.
Mamm Genome. 1996 Aug;7(8):580-5. doi: 10.1007/s003359900173.
The random amplified polymorphic DNA (RAPD) assay utilizes the polymerase chain reaction (PCR) and short primers of arbitrary nucleotide sequence to amplify DNA. In this study, the RAPD assay was used to identify and map polymorphic markers in the AgResearch International Mapping Flock (IMF) sheep pedigrees. Sires and dams of eight of the full-sib IMF pedigrees were screened with 131 different 10-mer oligonucleotide primers. An average of 85 RAPD polymorphisms was identified between each parental pair, and 53 markers were contributed to the AgResearch IMF collaboration. Forty-five of the RAPD markers were mapped in the AgResearch IMF genetic linkage map, and at least one marker was located on 17 of the 26 autosomes and both sex chromosomes. Three lines of evidence were used to check for the homology of scored polymorphisms in different pedigrees, pedigree evaluation, segregation analysis, and Southern blot analysis. These results demonstrate that the RAPD assay is a powerful approach for identifying polymorphisms that can be used as markers for constructing a sheep genetic linkage map.
随机扩增多态性DNA（RAPD）分析利用聚合酶链反应（PCR）和任意核苷酸序列的短引物来扩增DNA。在本研究中，RAPD分析用于鉴定和定位AgResearch国际作图群体（IMF）绵羊系谱中的多态性标记。使用131种不同的10聚体寡核苷酸引物对8个全同胞IMF系谱的父本和母本进行了筛选。每个亲本对之间平均鉴定出85个RAPD多态性，并且有53个标记用于AgResearch IMF合作项目。45个RAPD标记被定位到AgResearch IMF遗传连锁图谱中，并且在26条常染色体和两条性染色体中的17条上至少定位到了一个标记。使用了三条证据线来检查不同系谱中计分多态性的同源性，即系谱评估、分离分析和Southern印迹分析。这些结果表明，RAPD分析是一种强大的方法，可用于鉴定可作为构建绵羊遗传连锁图谱标记的多态性。
