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Combined DNA-RNA fluorescent in situ hybridization (FISH) to study X chromosome inactivation in differentiated female mouse embryonic stem cells.

TitleCombined DNA-RNA fluorescent in situ hybridization (FISH) to study X chromosome inactivation in differentiated female mouse embryonic stem cells.
Publication TypeJournal Article
Year of Publication2014
AuthorsBarakat TStefan, Gribnau J
JournalJ Vis Exp
Issue88
Date Published2014
ISSN1940-087X
KeywordsAnimals, Cell Differentiation, DNA, Embryonic Stem Cells, Female, In Situ Hybridization, Fluorescence, Mice, RNA, X Chromosome Inactivation
Abstract

Fluorescent in situ hybridization (FISH) is a molecular technique which enables the detection of nucleic acids in cells. DNA FISH is often used in cytogenetics and cancer diagnostics, and can detect aberrations of the genome, which often has important clinical implications. RNA FISH can be used to detect RNA molecules in cells and has provided important insights in regulation of gene expression. Combining DNA and RNA FISH within the same cell is technically challenging, as conditions suitable for DNA FISH might be too harsh for fragile, single stranded RNA molecules. We here present an easily applicable protocol which enables the combined, simultaneous detection of Xist RNA and DNA encoded by the X chromosomes. This combined DNA-RNA FISH protocol can likely be applied to other systems where both RNA and DNA need to be detected.

DOI10.3791/51628
Alternate JournalJ Vis Exp
PubMed ID24961515
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