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An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
Published on: May 23, 2018
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Complete sequence-based pathway analysis by differential on-chip DNA and RNA extraction from a single cell.
D van Strijp1, R C M Vulders1, N A Larsen2
1Philips Research Laboratories, High Tech Campus 11, 5656 AE, Eindhoven, The Netherlands.
Scientific Reports
|September 10, 2017
Summary
This study presents a novel microfluidic chip for simultaneous DNA and RNA extraction from single cells. This method aids in analyzing tumor pathways and mutations in individual cells, valuable for cancer research.
Area of Science:
- Biotechnology
- Genomics
- Molecular Biology
Background:
- Single-cell analysis is crucial for understanding cellular heterogeneity in diseases like cancer.
- Current methods for DNA and RNA extraction from single cells can be complex and lead to sample loss.
Purpose of the Study:
- To develop and validate an on-chip method for differential DNA and RNA extraction from a single cell.
- To enable simultaneous functional and mutational analysis of individual cells for cancer research.
Main Methods:
- Utilized a microfluidic chip with a two-stage lysis protocol for simultaneous DNA and RNA extraction.
- Applied a Bayesian computational pathway model to RNA sequences for pathway analysis.
- Analyzed DNA sequences from the same cell to identify pathway-driving mutations.
Main Results:
- Successfully demonstrated on-chip differential DNA and RNA extraction from single cells.
- Enabled direct use of whole extracts without washing steps, minimizing sample loss.
- Determined tumor-driving pathways and mutations in individual colorectal cancer cells.
Conclusions:
- The developed method allows for combined functional and mutational assessment of single cells.
- This approach holds significant value for dissecting tumor heterogeneity and analyzing circulating tumor cells.
