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单分子荧光原位杂交 (SM-FISH) 在小鼠卵母细胞中 Mrna 的定量和定位

JoVE Journal
Genética

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JoVE Journal Genética

Use of Single Molecule Fluorescent In Situ Hybridization (SM-FISH) to Quantify and Localize mRNAs in Murine Oocytes

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单分子荧光原位杂交 (SM-FISH) 在小鼠卵母细胞中 Mrna 的定量和定位

DOI:

10.3791/59414-v

•

08:18 min

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April 24, 2019

•

Kelsey R. Timme, Jennifer R. Wood

¹Department of Animal Science,University of Nebraska-Lincoln

Capítulos

00:04Título
00:38Collection of Ovulated Oocytes from Female Mice
02:06SM-FISH Staining of Oocytes
04:21Image Processing
06:21Results: mRNA Quantification Results using the Spot Finding and Tracking Program
07:45Conclusion

Summary

Tadução automática

为了可重复地计数单个卵母细胞中的 Mrna 数量, 对非粘附细胞的单分子 RNA 荧光原位杂交 (RNA-FISH) 进行了优化。卵母细胞被收集, 与转录特异性探针杂交, 并使用图像定量软件进行量化。

Tags

Single Molecule Fluorescent In Situ Hybridization (SM-FISH)MRNA Quantification MRNA Localization Murine Oocytes Nanog Pou5f1 DapB Oocyte Preparation Fixation Permeabilization Protease Treatment Probe Hybridization

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