为了应用 RFL P分析猪线粒体 DNA D loop的多态性。应用 PCR对西双版纳近交系小耳猪、广西巴马小型猪、贵州小型香猪、大约克夏猪、荣昌猪和长白猪血液总 DNA样品中线粒体 DNA控制区 (mt DNA D loop)进行扩增 ,2 3种限制性内切酶消化 ,...为了应用 RFL P分析猪线粒体 DNA D loop的多态性。应用 PCR对西双版纳近交系小耳猪、广西巴马小型猪、贵州小型香猪、大约克夏猪、荣昌猪和长白猪血液总 DNA样品中线粒体 DNA控制区 (mt DNA D loop)进行扩增 ,2 3种限制性内切酶消化 ,琼脂糖凝胶电泳检测。结果发现 mt DNA D loop扩增带清晰 ,与已知片段长度一致 ,RFL P分析未见长度多态。可见猪线粒体 DNA D loop酶切多态性贫乏。因此根据现用酶 ,应用mt DNA D loop PCR RFL P分析 。展开更多
Preparation of pure mitochondrial DNA (mtDNA) generally represents a key step in the field of mtDNA study. All cell membranes except the nuclear envelope were lysed in a buffer containing 1% Triton X-100. After proper...Preparation of pure mitochondrial DNA (mtDNA) generally represents a key step in the field of mtDNA study. All cell membranes except the nuclear envelope were lysed in a buffer containing 1% Triton X-100. After proper centrifugation and nuclei/cytoplasm isolation, mtDNA was obtained in the supernatant fraction. Preparation of mtDNA using this method is possible to prepare a mtDNA-rich, nDNA-free cellular fraction form tissues which is ready to be directly used. The method is simple and rapid.展开更多
文摘为了应用 RFL P分析猪线粒体 DNA D loop的多态性。应用 PCR对西双版纳近交系小耳猪、广西巴马小型猪、贵州小型香猪、大约克夏猪、荣昌猪和长白猪血液总 DNA样品中线粒体 DNA控制区 (mt DNA D loop)进行扩增 ,2 3种限制性内切酶消化 ,琼脂糖凝胶电泳检测。结果发现 mt DNA D loop扩增带清晰 ,与已知片段长度一致 ,RFL P分析未见长度多态。可见猪线粒体 DNA D loop酶切多态性贫乏。因此根据现用酶 ,应用mt DNA D loop PCR RFL P分析 。
文摘Preparation of pure mitochondrial DNA (mtDNA) generally represents a key step in the field of mtDNA study. All cell membranes except the nuclear envelope were lysed in a buffer containing 1% Triton X-100. After proper centrifugation and nuclei/cytoplasm isolation, mtDNA was obtained in the supernatant fraction. Preparation of mtDNA using this method is possible to prepare a mtDNA-rich, nDNA-free cellular fraction form tissues which is ready to be directly used. The method is simple and rapid.
