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Sequence specific sorting of DNA molecules with FACS using 3dPCR.
David J Sukovich1,2, Shea T Lance1,2,3, Adam R Abate1,2,3
1Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, USA.
Scientific Reports
|January 5, 2017
Summary
Three-dimensional digital PCR (3dPCR) sorts DNA molecules by sequence specificity. This method recovers long DNA targets from heterogeneous samples for deep sequencing, aiding rare mutation detection in cancer and pathogen identification.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Genetic heterogeneity poses challenges in biological system characterization.
- Modern sequencing instruments yield short DNA reads, limiting comprehensive analysis.
- Analyzing rare DNA sequences in mixed populations is technically difficult.
Purpose of the Study:
- To introduce a novel method for sequence-specific DNA molecule sorting.
- To enable the recovery of long DNA targets from heterogeneous samples.
- To facilitate deep sequencing of rare genetic targets.
Main Methods:
- Development of three-dimensional digital PCR (3dPCR).
- Sequence-specific sorting of DNA molecules.
- Recovery of long DNA targets for subsequent deep sequencing.
Main Results:
- 3dPCR effectively sorts DNA molecules based on specific sequences.
- The method allows for the enrichment of rare DNA targets from complex mixtures.
- Enables deep sequencing of previously inaccessible long DNA targets.
Conclusions:
- 3dPCR is a valuable tool for characterizing genetic heterogeneity.
- The method has applications in cancer mutation analysis and pathogen detection.
- 3dPCR overcomes limitations of short-read sequencing for rare target recovery.
