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mRNA analysis of single living cells

Analysis of specific gene expression in single living cells may become an important technique for cell biology. So far, no method has been available to detect mRNA in living cells without killing or destroying them. We have developed here a novel method to examine gene expression of living cells usi...

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Detalles Bibliográficos
Autores principales: Osada, Toshiya, Uehara, Hironori, Kim, Hyonchol, Ikai, Atsushi
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2003
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC151804/
https://www.ncbi.nlm.nih.gov/pubmed/12646067
http://dx.doi.org/10.1186/1477-3155-1-2
Descripción
Sumario:Analysis of specific gene expression in single living cells may become an important technique for cell biology. So far, no method has been available to detect mRNA in living cells without killing or destroying them. We have developed here a novel method to examine gene expression of living cells using an atomic force microscope (AFM). AFM tip was inserted into living cells to extract mRNAs. The obtained mRNAs were analyzed with RT-PCR, nested PCR, and quantitative PCR. This method enabled us to examine time-dependent gene expression of single living cells without serious damage to the cells.