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1.9K Views
•
06:15 min
July 6th, 2022
DOI :
10.3791/64060-v
Chapters
0:04
Introduction
0:47
Centrifugation of the Oocytes
1:51
Preparation of the Oocyte for Bisection
3:00
Bisection of the Oocytes
4:56
Results: Successful and Unsuccessful qPCR Results and Boxplot from Mitochondria-Reduced and Mitochondria-Dense Ooplasts
5:39
Conclusion
Transcript
该方案显着降低了卵母细胞中的线粒体DNA拷贝数。这可能有利于种内克隆。该方法将牛卵母细胞中的线粒体DNA拷贝数减少90%以上,从而降低克隆胚胎中线粒体异质体的发生率。
劳拉·亚当斯(Laura Adams)是我实验室的一名博士生,他将演示该程序。首先,使用移液管,从HSOF滴剂中收集选定的成熟卵母细胞,并将它们放入含有50微升HSOF CB溶液的1.5毫升管中。将卵母细胞以15, 000倍G离心12分钟。
在离心卵母细胞时,准备二分切板。为此,首先使用每滴20微升在
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Summary
在这里,我们提出了一种方案,以显着减少牛卵母细胞中的线粒体DNA拷贝数(P <0.0001)。该方法利用离心和二分切来大大减少卵母细胞线粒体,并且可以增加在重建的种间体细胞核移植胚胎中发育的机会。
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