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Allele-specific detection of single mRNA molecules in situ

We describe a method for fluorescent in situ identification of individual mRNA molecules, allowing quantitative and accurate measurements of allele-specific transcripts that differ by only a few nucleotides, in single cells. By using a combination of allele-specific and non-allele-specific probe lib...

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Detalles Bibliográficos
Autores principales: Hansen, Clinton H., van Oudenaarden, Alexander
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3789122/
https://www.ncbi.nlm.nih.gov/pubmed/23934076
http://dx.doi.org/10.1038/nmeth.2601
Descripción
Sumario:We describe a method for fluorescent in situ identification of individual mRNA molecules, allowing quantitative and accurate measurements of allele-specific transcripts that differ by only a few nucleotides, in single cells. By using a combination of allele-specific and non-allele-specific probe libraries, we achieve over 95% detection accuracy. We investigate the allele-specific stochastic expression of the pluripotency factor Nanog in murine embryonic stem cells.