摘要
本文详细报道了一个针对棉花特点的线粒体以及线粒体ＤＮＡ的抽提方法，即先用蔗糖Ｓｔｅｐ递度和ＤＮａｓｅＩ体外消化分离纯化线粒体，然后氯仿抽提纯化ＤＮＡ。实验过程中采用ＰＶＰ排除棉酚和丹宁的干扰，用ＤＩＥＣＡ抑制酚氧化酶活性，并对许多实验细节作了较大改进。得到的ＤＮＡ可满足酶切、ＰＣＲ等分子生物学分析。
A method suitable for extraction of cotton mitochondria and mitochondrial DNA(mtDNA)is reported in details.Mitochondria is separated by centrifugation in a sucrosestep density gradient and purified through DNase I digestion. mtDNA is extracted withchloroform. PVP and DIECA are used to bind phenolic compounds and danin and to inacti-vate phenol oxidases,respectively. Many operation steps are greatly improved in this pro-tocol.mtDNA prepared by this method for restriction analvsis,PCR and other molecularoperations were successfully used.
出处
《棉花学报》
CSCD
北大核心
1996年第6期316-317,共2页
Cotton Science
关键词
棉花
线粒体
线粒体DNA
Cotton mitochondria mitochondrial DNA(mtDNA).
