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Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay
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Journal JoVE Génétique
Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay

Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay

4,854 Views

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10:32 min

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February 03, 2022

DOI:

10.3791/63448-v

DOI:
10.3791/63448-v

•

10:32 min
February 03, 2022

•

28 Views
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1Department of Microbiology, Institute of Biomedical Sciences,University of São Paulo, 2Division of Oncology, Department of Internal Medicine,Washington University in St. Louis

Summary

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Here we describe two modifications of the DNA fiber assay to investigate single-stranded DNA gaps in replicating DNA after lesion induction. The S1 fiber assay enables the detection of post-replicative gaps using the ssDNA-specific S1 endonuclease, while the gap-filling assay allows visualization and quantification of gap repair.

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Cite this Article

Martins, D. J., Tirman, S., Quinet, A., Menck, C. F. M. Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay. J. Vis. Exp. (180), e63448, doi:10.3791/63448 (2022).

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