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CRISPR-based DNA and RNA detection with liquid phase separation

View ORCID ProfileWillem Kasper Spoelstra, Jeroen M. Jacques, View ORCID ProfileFranklin L. Nobrega, Anna C. Haagsma, Marileen Dogterom, View ORCID ProfileTimon Idema, View ORCID ProfileStan J. J. Brouns, View ORCID ProfileLouis Reese
doi: https://doi.org/10.1101/471482
Willem Kasper Spoelstra
1Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands
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Jeroen M. Jacques
1Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands
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Franklin L. Nobrega
1Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands
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Anna C. Haagsma
1Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands
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Marileen Dogterom
1Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands
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Timon Idema
1Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands
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Stan J. J. Brouns
1Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands
2Laboratory of Microbiology, Wageningen University, Stippeneng 4, 6708 WE Wageningen, The Netherlands
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  • For correspondence: stanbrouns@gmail.com
Louis Reese
1Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands
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Abstract

The RNA-guided CRISPR endonucleases Cas12a and Cas13a allow detection of specific DNA or RNA sequences in molecular diagnostics applications. Here, we present a novel approach that facilitates sequence detection even with the unaided eye, through solution turbidity arising from liquid-liquid phase separation of nucleic acids and positively charged polyelectrolytes. The simplicity of combining CRISPR-nuclease activity with macroscopic phase separation allows for easy and low-cost detection of specific nucleic acids in a wide range of applications.

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Posted November 20, 2018.
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CRISPR-based DNA and RNA detection with liquid phase separation
Willem Kasper Spoelstra, Jeroen M. Jacques, Franklin L. Nobrega, Anna C. Haagsma, Marileen Dogterom, Timon Idema, Stan J. J. Brouns, Louis Reese
bioRxiv 471482; doi: https://doi.org/10.1101/471482
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CRISPR-based DNA and RNA detection with liquid phase separation
Willem Kasper Spoelstra, Jeroen M. Jacques, Franklin L. Nobrega, Anna C. Haagsma, Marileen Dogterom, Timon Idema, Stan J. J. Brouns, Louis Reese
bioRxiv 471482; doi: https://doi.org/10.1101/471482

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