Single cell sequencing
The disclosure provides methods and systems of analyzing single cells by simultaneously separating cells into monodisperse droplets and tagging each nucleic acid molecule from the cells with barcodes unique to each droplet. The methods and systems combine template particles with a plurality of single cells in a tube, generate in the tube monodispersed droplets encapsulating a single one of the template particles and a single one of the single cells, release nucleic acid molecules from the single cells and provide each nucleic acid molecule with a barcode unique to the respective droplet. The nucleic acid molecules can then be analyzed by any known method, for example by sequencing the nucleic acid molecules.
1. A method for single cell analysis, the method comprising:
combining a plurality of template particles, each comprising a unique barcode, and single cells;
generating a plurality of monodispersed droplets simultaneously that encapsulate a single template particle and a single cell;
releasing nucleic acid molecules from the single cells; and
analyzing the nucleic acid molecules.
2. The method of claim 1 , wherein the nucleic acid molecules are DNA or RNA molecules.
3. The method of claim 1 , wherein the template particles comprise one selected from the group consisting of agarose, alginate, polyethylene glycol, polyacrylamide, acrylate, acrylamide, and bis-acrylamide.
4. The method of claim 1 , wherein the template particles are spherical.
5. The method of claim 1 , wherein the monodisperse droplets are deposited in a tube.
6. The method of claim 1 , wherein the template particles are dehydrated prior to said combining step.
7. The method of claim 1 , wherein said single cells are obtained from a tissue or body fluid sample.
8. The method of claim 7 , wherein the number of cells to be analyzed is 100,000 cells.
9. The method of claim 1 , further comprising associating the nucleic acid molecules with cells from which they were obtained.
10. The method of claim 1 , further comprising analyzing said nucleic acids for identification of a mutation.
11. The method of claim 10 , wherein the mutation constitutes a cancer genotype.
12. The method of claim 11 , further comprising staging cancer based on said genotype.
13. The method of claim 11 , further comprising prescribing a treatment protocol.
14. The method of claim 1 , wherein said analyzing step comprises PCR.
15. The method of claim 1 , wherein the template particles are faceted.
16. The method of claim 1 , wherein the template particles comprise internal compartments.
17. The method of claim 16 , wherein the template particles are capable of triggered release of reagents from said internal compartments.
18. The method of claim 3 , wherein said RNA is an mRNA.