Solution-phase indexing by kinetic confinement enables rapid, simple, and instrument-free single cell transcriptional profiling
Marafini, P.; Smith, D. G.; Lamstaes, A. R.; Contreras, R. E.; Williams, I.; West, I.; Ambridge, O.; Sanders-Brown, V.; Intaite, E.; Hii, C. Y.; Hume, B. C.; Munagala, U.; Plumbly, W.; Brown, F. L.; Shlyakhtina, Y.; Woods, L.; Bibby, J. A.; Williams, L.; Yang, J. H.; Steffy, B.; Zawada, L.; Harger, J. W.; McKenzie, D.; Laing, A. G.; Stubbington, M. J.; Edelman, L. B.
Show abstract
Existing tools for single cell genomics require complex physical frameworks for the indexing of cellular nucleic acids, including proprietary instrumentation, droplet emulsions, and laborious combinatorial indexing schemes. The complexity and cost of these tools significantly constrains the use of single cell technologies across basic and translational research. Here, we describe an instrument-free method that uses novel, bifunctional indexing reagents to deliver index sequences directly to single cells followed by a biophysical process known as Kinetic Confinement to perform high-fidelity indexing of target molecules across thousands of single cells simultaneously in single-tube, solution-phase reactions. Kinetic Confinement enables simple, fast, and flexible single cell experiments, and allows straightforward scaling to very large sample numbers. We anticipate that assays based on Kinetic Confinement will significantly expand the scope, use, and impact of single cell analysis across fundamental and applied research, as well as within therapeutic development and ultimately applied clinical diagnostics.
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