Detection of RNA-DNA hybrids by immunostaining in meiotic nuclei of Saccharomyces cerevisiae

RNA transcripts can anneal with template DNA strands to form RNA-DNA hybrids (or R-loops if the non-template DNA strands exist), which play a variety of roles in many physiological processes. Here, we provide an accessible and reproducible approach for immunofluorescent staining of RNA-DNA hybrids w...

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Detalles Bibliográficos
Autores principales: Yang, Xuan, Zhang, Liangran
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9035804/
https://www.ncbi.nlm.nih.gov/pubmed/35479113
http://dx.doi.org/10.1016/j.xpro.2022.101325
Descripción
Sumario:RNA transcripts can anneal with template DNA strands to form RNA-DNA hybrids (or R-loops if the non-template DNA strands exist), which play a variety of roles in many physiological processes. Here, we provide an accessible and reproducible approach for immunofluorescent staining of RNA-DNA hybrids with the S9.6 antibody in spread meiotic nuclei of Saccharomyces cerevisiae. This protocol allows the examination of RNA-DNA hybrids as clearly distinguishable foci and the colocalizations of RNA-DNA hybrids with other proteins. For complete details on the use and execution of this protocol, please refer to Yang et al. (2021).

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