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Sci Rep ; 7(1): 11030, 2017 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-28887473

RESUMO

We demonstrate on-chip, differential DNA and RNA extraction from a single cell using a microfluidic chip and a two-stage lysis protocol. This method enables direct use of the whole extract, without additional washing steps, reducing sample loss. Using this method, the tumor driving pathway in individual cells from a colorectal cancer cell line was determined by applying a Bayesian computational pathway model to sequences obtained from the RNA fraction of a single cell and, the mutations driving the pathway were determined by analyzing sequences obtained from the DNA fraction of the same single cell. This combined functional and mutational pathway assessment of a single cell could be of significant value for dissecting cellular heterogeneity in tumors and analyzing single circulating tumor cells.


Assuntos
DNA/isolamento & purificação , Redes Reguladoras de Genes , Microfluídica/métodos , RNA/isolamento & purificação , Análise de Célula Única/métodos , Linhagem Celular Tumoral , Neoplasias Colorretais/patologia , Misturas Complexas/análise , Misturas Complexas/isolamento & purificação , DNA/análise , Humanos , RNA/análise
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