1. Complete sequence-based pathway analysis by differential on-chip DNA and RNA extraction from a single cell
- Author
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Roland C. M. Vulders, Loïc Baerlocher, Nicholas A. Larsen, Rodolphe Marie, Thomas Steen Hansen, Marie Pødenphant, M. A. van Driel, P. J. van der Zaag, Kalim U. Mir, Julien Schira, Tom Olesen, Wim Verhaegh, D. van Strijp, and Anders Kristensen
- Subjects
0301 basic medicine ,Lysis ,Microfluidics ,Cell ,lcsh:Medicine ,Complex Mixtures ,Biology ,Article ,03 medical and health sciences ,Complete sequence ,chemistry.chemical_compound ,Circulating tumor cell ,Cell Line, Tumor ,medicine ,Humans ,Gene Regulatory Networks ,lcsh:Science ,Multidisciplinary ,lcsh:R ,RNA ,DNA ,Pathway analysis ,Molecular biology ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,lcsh:Q ,RNA extraction ,Single-Cell Analysis ,Colorectal Neoplasms - Abstract
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.
- Published
- 2017
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