琼脂糖和聚丙烯酰胺凝胶电泳技术检测小麦基因组DNA RAPD扩增产物的方法学比较

通过20%(wv)的琼脂糖凝胶和5%(wv)的聚丙烯酰胺凝胶电泳对小麦白粉病抗、感特性品种基因组DNA的RAPD检测结果表明:5%聚丙烯酰胺凝胶对线性DNA分子(01～20kb)和长度相差100bp以下的DNA分子的分离较20%的琼脂糖凝胶电泳效果好。因此,我们研究出了一项利用聚丙烯酰胺凝胶电泳检测小麦白粉病抗、感特性的新技术,在工作中建立了一种适合于检测小麦基因组DNA结构差异的电泳方法。该方法主要包括:(1)丙烯酰胺和亚甲基双丙烯酰胺的新配比;(2)分离DNA片段的最佳凝胶浓度;(3)电泳条件;(4)脱色、漂洗、银染、显色过程。实验发现,该技术对于小麦白粉病抗、感特性检测中的小片段和长度相差100bp以下的线性DNAPCR扩增结果的分辨效果较好。应用该技术在抗感品种间已经发现了DNA水平上的差异。

The Comparison of RAPD Detecting Results of Breeds for Resistant or Sensitive Species of Powdery Mildew in Wheat Using Ago-Gel and PAG Electrophoretic Technologies

w/v)Ago-gel as maximum gel-density is inferior to 5%(w/v)PAG for some small segments and some DNA molecules which length differ within 100bp by way of comparing of RAPD detecting results of breeds for resistant or sensitive species of powdery mildew in wheat.Consequently an improved PAGE technology suitable for detecting results of breeds for resistant or sensitive species of powdery mildew in wheat was developed.The technology characterized by(1)a new ration of Acr and Bis in gel stock solution,(2)a best gel density is perfect for separating DNA segments,(3)the better electrophoretic condition,(4)the procedure of discoloring,bleaching,silver dying and developing.Test discovered that the technology distinguished effect is good for some small segments and some DNA molecules which length differ within 100bp.The technology has discovered DNA difference in breeds for resistant or sensitive species of powdery mildew in wheat. ;