延边地区圈养黑熊基因DNA中部分微卫星位点的探讨

本研究采用分光光度法及琼脂糖凝胶电泳比较从黑熊组织、血液以及毛发中提取的黑熊基因组DNA,发现提取的DNA浓度及纯度均可满足后续试验的需要。其中毛发DNA提取方法具有高灵敏度、操作简单、非损伤等特点,有助于开展珍稀濒危物种和性情凶猛动物的遗传特性研究。本文还根据文献中黑熊的11个微卫星位点设计并合成引物,进行了不同引物PCR最佳反应条件的筛选。对黑熊DNA基因组PCR扩增结果发现其中10对引物能扩增出较明显的扩增带,为后续的黑熊亲子鉴定及遗传特异性分析实验提供了有价值的资料,为其亲权概率的准确性提供了良好的参考依据。

In this study,we compared the quality of genome DNA extracted from the tissue or blood or hair of black bears using spectrophotgmeter and agarose gel eleetrophoresis and found that the concentration and purity of genome DNA we obtained can all meet the needs of the subsequent experiments. Among three resources of genomic DNA, the hair extraction method has many benefits like high sensitivity, simple operation, non-damage etc, may solve the difficult samplings from the rare and endangered animal species or the temperament fierce animals, and may help the investigators to conduct many important genomic studies of them. We investigated 11 microsatellite loci of the black bear,designed and synthesized every set of primers depending on the sequences of each microsatellite, finally screened the best reaction conditions for polymerase chain reaction （PCR） of every microsatellite. The PCR results showed ~hat 10 in 11 microsatellites mentioned above have displayed relatively clear PCR bands using synthesized primer set respectively. These 10 microsatellites have high values in developing accurate paternity test in the black bear species, and will be a good reference basis to the paternity rates calculations.