核酸凝胶电泳
核酸凝胶电泳是根据DNA和RNA片段的大小及带电量对其进行分离、辨别和提纯的一种分子生物学技术。该技术利用外加电场使带负电荷的核酸分子穿过琼脂糖或者聚丙烯酰胺凝胶基质,从而分离出DNA和RNA核酸分子。
其中凝胶基质可起到筛选作用,因为小分子能比大分子更快地穿过凝胶基质孔隙。
- 一般来说,琼脂糖凝胶常用于DNA和RNA的电泳实验中,结合标准的电泳技术,可以分离50到50000碱基对的DNA片段。
- 还有一种分离范围较小的聚丙烯酰胺凝胶,可分离小于500个碱基对的DNA片段,但分辨率高。
- 两种凝胶都可用于凝胶阻滞分析(EMSA)实验,来表征蛋白和核酸的相互作用。
经凝胶电泳分离后,DNA和RNA分子可通过染色或者紫外光照来观察、从凝胶中提取和纯化或者通过转膜进行印迹分析。这些都是克隆、聚合酶链式反应、质谱、高通量测序、RNA和DNA印迹杂交等工作中常用的制备技术。
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