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JoVE Journal Genetics
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

Article DOI: 10.3791/63448-v • 10:32 min • February 3rd, 2022
February 3rd, 2022 •
*1, *2, 2, 1
1Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, 2Division of Oncology, Department of Internal Medicine, Washington University in St. Louis
* These authors contributed equally

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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.

Tags

DNA Fiber Assay Post-replicative Gaps DNA Lesion S1 Nuclease DNA Repair Replication Stress Response Genome Integrity Controls IDU Media UV-C Irradiation
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