摘要
建立灵敏、可靠的大鼠内耳膜迷路线粒体ＤＮＡ（ｍｔＤＮＡ）提取和检测方法。方法：结合ＰＣＲ技术扩增ｍｔＤＮＡ编码ＮＤ１－１６ ＳｒＲＮＡ基因的 ６０１ ｂｐ片段，检测大鼠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只大鼠的双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ＮＤ １－１６ ＳｒＲＮＡ基因的 ６０１ ｈｐ片段；而采用 Ｅｄｒｉｓ ｍｔＤＮＡ提取方法，需用 ６侧听泡内耳膜迷路作为一个样品方可获得可靠的阳性结果。结论：采用Ｓｅｉｄｍａｎ方法提取内耳膜迷路线粒体ＤＮＡ，结合ＰＣＲ技术扩增ｍｔＤＮＡ高度保守基因片段，作为总ＤＮＡ中含有被ＰＣＲ扩增量ｍｔＤＮＡ存在指标，可应用于内耳膜迷路ｍｔＤＮＡ突变的研究。
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.
出处
《临床耳鼻咽喉科杂志》
CSCD
1999年第4期176-177,共2页
Journal of Clinical Otorhinolaryngology
关键词
聚合酶链反应
内耳膜迷路
线粒体DNA
PCR Inner ear
Membranous labyrinth
Mitochondria
DNA detection
