A DNA-fiber protocol for single molecule analysis of telomere (SMAT) length and extension events in cancer cells

Alternative lengthening of telomeres (ALT) is a homologous recombination-based telomere maintenance mechanism. It is active in approximately 10–15% of cancers. We present a DNA-fiber protocol, combining YOYO-1 staining of genomic DNA, telomere fluorescence in situ hybridization (FISH), and EdU label...

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Detalles Bibliográficos
Autores principales: Lu, Robert, Allen, Joshua A.M., Galaviz, Pablo, Pickett, Hilda A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8899046/
https://www.ncbi.nlm.nih.gov/pubmed/35265860
http://dx.doi.org/10.1016/j.xpro.2022.101212
Descripción
Sumario:Alternative lengthening of telomeres (ALT) is a homologous recombination-based telomere maintenance mechanism. It is active in approximately 10–15% of cancers. We present a DNA-fiber protocol, combining YOYO-1 staining of genomic DNA, telomere fluorescence in situ hybridization (FISH), and EdU labeling of nascent DNA, to measure telomere extension events in ALT cancer cells. The protocol can be used to delineate ALT-mediated telomere extension. For complete details on the use and execution of this protocol, please refer to Barroso-Gonzalez et al. (2021).

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