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Forbedret Cross Linking Immunoprecipitation (eCLIP) metode til effektiv identifikation af protein-bundet RNA i Mouse testis

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JoVE Journal Biology

Enhanced Crosslinking Immunoprecipitation (eCLIP) Method for Efficient Identification of Protein-bound RNA in Mouse Testis

Forbedret Cross Linking Immunoprecipitation (eCLIP) metode til effektiv identifikation af protein-bundet RNA i Mouse testis

DOI:

10.3791/59681-v

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10:31 min

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May 10, 2019

•

Qiushi Xu*¹, Caifeng Wang*¹, Li Ling*¹, Qiuling Yue*¹, Mengrou Liu, Shuya Zhang, Kaiqiang Fu, Lan Ye, Ke Zheng

¹State Key Laboratory of Reproductive Medicine,Nanjing Medical University

Chapters

00:04Title
01:14Tissue Harvesting and UV Crosslinking
02:37Beads Preparation
03:20Tissue Lysis and Partial RNA Digestion
04:30Immunoprecipitation
05:09RNA Adapter Ligation to RNA 3’ Ends
06:48SDS-PAGE and Membrane Transfer
08:24Results: Analysis of the eCLIP Method
10:08Conclusion

Summary

Automatic Translation

Her præsenterer vi en eCLIP-protokol for at bestemme store RNA-mål for RBP-kandidater i testikler.

Tags

Keywords: ECLIP Protein-bound RNA Mouse Testis RNA-binding Proteins Spermatogenesis Non-radioactive Size-matched Input Small-scale Sequencing Crosslinking Immunoprecipitation Magnetic Beads

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