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以甜樱桃‘红灯’为试材,应用选择性扩增微卫星(SAM)法分离、克隆了100个SSR序列,其中81个非重复,可用。加上搜索数据库所获得的1个SSR序列,一共82个序列用于特异引物的设计。仅从69个序列的77个基因座设计出特异引物。合成38对特异引物,对其中的36个基因座进行检测。其中19对引物扩增出相应大小的片段,另外8对引物扩增出非预期片段。最后,以27个甜樱桃种质的基因组DNA为模板,从27对可扩增出带的引物中,筛选出多态性引物24对,获得了24个甜樱桃基因座特异性SSR标记。
Using sweet cherry ’red light’ as test material, 100 SSR sequences were cloned by selective amplification of microsatellite (SAM) method, of which 81 were non-repetitive and available. Add one SSR sequence obtained from the search database and a total of 82 sequences are used for the design of the specific primers. Specific primers were designed from 77 loci of 69 sequences. 38 pairs of specific primers were synthesized and 36 loci were detected. Among them, 19 pairs of primers amplified fragments of the corresponding size, and another 8 pairs of primers amplified unidentified fragments. Finally, 24 pairs of polymorphic primers were screened out from 27 pairs of primers, using 27 genomic DNAs from sweet cherry germplasm as template. 24 sweet cherry loci-specific SSR markers were obtained.