PCR技术对大鼠内耳膜迷路线粒体DNA的检测

目的：建立灵敏、可靠的大鼠内耳膜迷路线粒体ＤＮＡ提取和检测方法。方法：结合ＰＣＲ技术扩增ｍｔＤＮＡ编码ＮＤ１－１６ＳｒＲＮＡ基因的６０１ｂｐ片段，检测大听泡内耳膜迷路线粒体ＤＮＡ方法，并对两种ｍｔＤＮＡ获取方法进行比较。结果：采用Ｓｅｉｄｍａｎ的方法，将１只大鼠的以侧听泡内耳膜迷路作为一个样品，检测１０个样品均成功扩增出编码ＮＤ１－１６ＳｒＲＮＡ基因的６０１ｂｐ片段；而采用ＥｄｒｉｓｍｔＤＮＡ提取

Use PCR technique to detect mitochondrial DNA in rat membranous labyrinth

Objective:To establish a sensitive and reliable method of mtDNA detection in the membranous labyrinth of rat inner ear. Method: A mtDNA segment of 601 hp containing the genes which encode NDI subunit 165 rRNA in rat inner ear membranous labyrinth was detected by PCR method. The method is compared with Edris's mtDNA isolation method. Result=A 601 hp PCR product of mtDNA in the membranous labyrinth of rat ear inner can be obtained by the two methods. But the requirement of the sample quantities was different for the two methods. The extraction of mtDNA need two of inner ears membranous labyrinth to get reliably result by the method adapted from Seidman. However, the mtDNA extraction procedure from Edris's method need six of inner ears of the in order to get reliably result. Conclusion:The method of mtDNA isolation.amplification and detection membranous labyrinth of rat inner ear by PCR method adapted from seidman is more sersitive.