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Please note that all translations are automatically generated. Click here for the English version.
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02:46 min
March 17th, 2023
DOI :
10.3791/200013-v
Transcript
一旦在第7天获得约2500万个线粒体供体细胞,在50毫升管中收获指数生长的细胞,并通过在室温和520g下离心五分钟来收集它们。用冷磷酸盐缓冲盐水洗涤细胞,并在室温下以520g离心五分钟。现在,使用冷试剂在四摄氏度下进行整个线粒体提取,并将细胞或线粒体保持在冰上。
第三次离心后,使用与真空泵耦合的玻璃移液管抽吸弃上清液,并将填充的细胞重悬于等于细胞沉淀体积七倍的低渗缓冲液中。然后将细胞悬液转移到均质管中,并通过在冰上孵育两分钟让细胞膨胀。通过在均质器中执行 8 到 10 次冲程来
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