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

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8.3K Views

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10:10 min

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May 28th, 2017

DOI :

10.3791/55919-v

May 28th, 2017

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Oriana Romano¹^,², Ingrid Cifola³, Valentina Poletti⁴, Marco Severgnini³, Clelia Peano³, Gianluca De Bellis³, Fulvio Mavilio¹^,⁴, Annarita Miccio¹^,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

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Keywords 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

Rozdziały w tym wideo

0:05

Title

0:45

MLV Transduction of Human Cells

1:41

Restriction Enzyme Digestion and Linker Ligation

3:05

First-round Linker-mediated PCR

3:43

Second Round Linker-mediated PCR

5:17

Library Preparation and Dilution for Massive Sequencing of MLV Integration Sites

7:53

Results: Mapping MLV Integration into Epigenetically-defined Regulatory Regions

9:23

Conclusion

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