Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells

Summary: Molecular cloning of BRD4-L is a challenging technique, because the DNA insert is formed by a long, GC-rich sequence, which folds into secondary structures. The present protocol defines a specific strategy to amplify BRD4-L, followed by the successful cloning of the gene into an overexpress...

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Bibliographic Details
Main Authors: Ana Luiza Drumond-Bock, Magdalena Cybula, Luyao Wang, Lin Wang, Magdalena Bieniasz
Format: Article
Language:English
Published: Elsevier 2022-12-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166722006657
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Summary:Summary: Molecular cloning of BRD4-L is a challenging technique, because the DNA insert is formed by a long, GC-rich sequence, which folds into secondary structures. The present protocol defines a specific strategy to amplify BRD4-L, followed by the successful cloning of the gene into an overexpression vector. Since there are no existing protocols nor commercially available plasmids, this work provides a useful tool for studies involving molecular cloning of BRD4-L and could potentially be applied to other challenging genes. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667