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利用下一代序列染色体构象捕获 (4 c 序列) 的高通量识别基因调控数列

JoVE Journal
Genetics

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JoVE Journal Genetics

High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)

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利用下一代序列染色体构象捕获 (4 c 序列) 的高通量识别基因调控数列

DOI:

10.3791/58030-v

•

09:06 min

•

October 05, 2018

•

Erin A. Brettmann, Inez Y. Oh, Cristina de Guzman Strong

¹Division of Dermatology, Center for Pharmacogenomics, Center for the Study of Itch, Department of Medicine,Washington University School of Medicine

Chapters

00:04Title
00:45Formaldehyde Cross-linking and Cell Lysis
01:59First Restriction Digestion and First Ligation
05:07Cross-linking Reversion, Chromatin Isolation, and Further Analyses
07:42Results: Representative Genomic Track of 4C-seq Read Coverage
08:29Conclusion

Summary

Automatic Translation

基因和调控元素之间的物理相互作用的识别是具有挑战性的, 但已被染色体构象捕获方法所促进。这种对 4 c 序列协议的修改可以减少 pcr 模板的过度放大, 并通过加入限制酶消化步骤来最大限度地提高 mappability 的读数。

Tags

4C-seq Chromatin Interactions Gene Regulatory Sequences Next-generation Sequencing Transcriptional Regulation Promoter-enhancer Interactions Cross-linking Restriction Digestion Cell Lysis

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