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High-Throughput Quantification of the Synthesis and Distribution of mtDNA
May 05, 2023
基于图像的高通量线粒体DNA合成和分布定量
01:31 min
•
用于线粒体DNA合成的siRNA混合物的制备
00:45 min
•
细胞转染和5-溴-2'-脱氧尿苷(BrdU)在HeLa细胞中的掺入
01:19 min
•
HeLa细胞中线粒体DNA的5-溴-2'-脱氧尿苷(BrdU)标记
01:44 min
•
HeLa细胞固定与Hoechst 33342和阻断
01:05 min
•
向转染的HeLa细胞中添加一抗和二抗以监测mtDNA分布
02:04 min
•
定量图像分析,用于测量转染HeLa细胞中的mtDNA和基因表达水平
03:02 min
•
Lukasz S. Borowski
2
,
Karolina Kasztelan
3
,
Jolanta Czerwinska-Kostrzewska
2
,
Roman J. Szczesny
1
Faculty of Biology, Institute of Genetics and Biotechnology
,
University of Warsaw
,
2
Institute of Biochemistry and Biophysics
,
Polish Academy of Sciences
,
3
International Institute of Molecular and Cell Biology in Warsaw
Summary
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English (Original)
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描述了使用多孔板格式和自动免疫荧光成像来检测和量化mtDNA合成和分布的细胞中线粒体DNA(mtDNA)代谢动力学的程序。这可以进一步用于研究各种抑制剂,细胞应激和基因沉默对mtDNA代谢的影响。
Article