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Journal
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Biology
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Enhanced Crosslinking Immunoprecipitation (eCLIP) Method for Efficient Identification of Protein-bound RNA in Mouse Testis
JoVE Journal
Biology
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JoVE Journal
Biology
Enhanced Crosslinking Immunoprecipitation (eCLIP) Method for Efficient Identification of Protein-bound RNA in Mouse Testis
Enhanced Crosslinking Immunoprecipitation (eCLIP) Method for Efficient Identification of Protein-bound RNA in Mouse Testis
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
1
State Key Laboratory of Reproductive Medicine
,
Nanjing Medical University
Chapters
00:04
Title
01:14
Tissue Harvesting and UV Crosslinking
02:37
Beads Preparation
03:20
Tissue Lysis and Partial RNA Digestion
04:30
Immunoprecipitation
05:09
RNA Adapter Ligation to RNA 3’ Ends
06:48
SDS-PAGE and Membrane Transfer
08:24
Results: Analysis of the eCLIP Method
10:08
Conclusion
Summary
Automatic Translation
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Automatic Translation
Here, we present an eCLIP protocol to determine major RNA targets of RBP candidates in testis.
Tags
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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