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增强交联免疫沉淀 (eCLIP) 方法, 有效鉴定小鼠睾丸蛋白质结合 rna

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

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增强交联免疫沉淀 (eCLIP) 方法, 有效鉴定小鼠睾丸蛋白质结合 rna

DOI:

10.3791/59681-v

•

10:31 min

•

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

자동 번역

在这里, 我们提出了一个 eCLIP 协议, 以确定主要的 RBP 候选在睾丸的 rna 目标。

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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