摘要
将微卫星探针5′端生物素化后与链亲和素磁珠特异结合,用磁珠和探针的结合物与两端连接已知序列人工接头的中国李品种小黄李(Prunus salicinacv.Xiaohuangli)基因组DNA酶切片段杂交,以此杂交片段为模板用人工接头序列为引物进行PCR扩增,根据PCR产物测序结果设计引物作为微卫星DNA的标记引物.结果在随机挑选的36个克隆进行菌落PCR检测时,从31个阳性克隆中挑选18个克隆进行测序后获得了12条特异序列,设计的8对SSR引物均在5个中国李受试品种上获得了预期的扩增产物,其中4对引物在受试品种上表现出多态性.
The micro-satellite probe was combined with biotin at its 5＇-end,and then crossed with genomic DNA fragments of ＇ Xiaohuangli＇ （Prunus salicina） produced in the digestion with Hind Ⅲ and EcoR Ⅰ to which the adapters with known sequences had been attached at their two ends;The crossed fragments were used as the template to conduct PCR amplification with the adapter sequences as the primers and the primers were designed according to the sequenced PCR products as the micro-satellite DNA markers. While these primers were used to test 36 clones by PCR,18 clones screened from 31 positive clones were sequenced and then 12 specific sequences were obtained;with the eight pairs of the designed SSR primers the expected amplification products were obtained from the five tested varieties of P. salicina and four of the eight pairs appeared polymorphic in the tested varieties.
出处
《西北植物学报》
CAS
CSCD
北大核心
2006年第7期1320-1325,共6页
Acta Botanica Boreali-Occidentalia Sinica
