甜菜SSR扩增产物不同检测方法的比较研究

利用16对甜菜SSR引物对12个不同的甜菜品种进行扩增,分别采用8%的非变性聚丙烯酰胺凝胶电泳、3%的Metaphor琼脂糖电泳以及3%的普通琼脂糖电泳对SSR扩增产物进行检测,比较不同检测结果的差异性。结果表明,8%的非变性聚丙烯凝胶分辨细小条带的差异远高于3%的Metaphor琼脂糖和普通琼脂糖,而Metaphor琼脂糖的的分辨率高于普通琼脂糖,但对于条带较少的引物,三种凝胶未显现出差异；对于条带数较多的引物,如果条带比较集中,三种检测方法差异显著,如果条带比较分散,三种凝胶之间的差异较小。由于Metaphor琼脂糖价格偏高、胶软不易操作且分辨率低于聚丙烯酰胺凝胶,因此不建议使用。而8%的聚丙烯酰胺凝胶则适合于指纹图谱的构建以及遗传多样性分析等等,普通琼脂糖可以应用在品种纯度的鉴定。

Methods to Detect the SSR Amplified Product in Sugar Beet: A Comparative Study

Using 16 pairs of sugar beet SSR primers to amplify 12 different sugar beet varieties, 8% nondenaturant polyacrylamide gel electrophoresis, 3% Metaphor agarose electrophoresis and 3% agarose gel electrophoresis were adopted to test the products of SSR amplification respectively and the differences between the test results were compared. The results demonstrated that: the resolution capacity of 8% non-denaturant polyacrylamide gel on the difference in tiny bands was much higher than that of the 3% Metaphor agarose and the 3% agarose gel; the resolution ratio of Metaphor agarose was higher than that of the agarose gel. However, for primers with fewer bands, the 3 gels showed no difference; for primers with more bands, if the bands were concentrated, the 3 detection methods would produce significant difference; if the bands were scattered, the 3 gels would only present small difference. Since Metaphor agarose has higher prices, softer gel (harder to handle) and lower resolution ratio than polyacrylamide gel, it is not recommended. 8% polyacrylamide gel is suitable for fingerprint construction and genetic diversity analysis while agarose gel is applicable to identifying the purity of variety.