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逆转录病毒扫描：映射MLV集成站点以定义细胞特异性监管区域

JoVE Journal
Genetica

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JoVE Journal Genetica

Retroviral Scanning: Mapping MLV Integration Sites to Define Cell-specific Regulatory Regions

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逆转录病毒扫描：映射MLV集成站点以定义细胞特异性监管区域

DOI:

10.3791/55919-v

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10:10 min

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May 28, 2017

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Oriana Romano², Ingrid Cifola, Valentina Poletti, Marco Severgnini, Clelia Peano, Gianluca De Bellis, Fulvio Mavilio⁴, Annarita Miccio^2,4,5

¹Center for Genome Research, Department of Life Sciences,University of Modena and Reggio Emilia, ²Laboratory of Chromatin and Gene Regulation During Development,Imagine Institute, ³Institute for Biomedical Technologies,CNR, ⁴Généthon, ⁵Sorbonne Paris Cité – Université Paris Descartes

Capitoli

00:05Titolo
00:45MLV Transduction of Human Cells
01:41Restriction Enzyme Digestion and Linker Ligation
03:05First-round Linker-mediated PCR
03:43Second Round Linker-mediated PCR
05:17Library Preparation and Dilution for Massive Sequencing of MLV Integration Sites
07:53Results: Mapping MLV Integration into Epigenetically-defined Regulatory Regions
09:23Conclusion

Summary

Traduzione automatica

在这里，我们描述了人类细胞中基于Moloney小鼠白血病病毒的逆转录病毒载体的整合位点全基因组图谱的方案。

Tags

Retroviral Scanning MLV Integration Sites Cell-specific Regulatory Regions Gene Therapy Gene Regulation MLV-derived Retroviral Vector EGFP Reporter Gene Flow Cytometric Analysis Genomic DNA Preparation Restriction Enzyme Digestion Linker Ligation First-round PCR Second-round PCR

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