Partitioning and processing of analytes and other species
The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing. Such polynucleotide processing may be useful for a variety of applications, including polynucleotide sequencing.
1. A system, comprising:
a plurality of capsules; and
a plurality of partitions,
wherein a partition of said plurality of partitions comprises (i) a first species and (ii) a capsule of said plurality of capsules, wherein said capsule comprises a second species, wherein said capsule is configured to release said second species into said partition upon application of a stimulus, and wherein said stimulus comprises one or more members selected from the group consisting of a change in ion concentration, a reducing agent, a light stimulus, an electrical stimulus, and a magnetic stimulus.
2. The system of claim 1 , wherein at least one of said first species and said second species is selected from the group consisting of a reagent and an analyte.
3. The system of claim 2 , wherein said reagent comprises one or more members selected from the group consisting of a protein, a polynucleotide, an enzyme, an antibody, a barcode, an adapter, a buffer, a small molecule, a detergent, a dye, and a polymer.
4. The system of claim 3 , wherein said enzyme comprises a proteinase, a restriction enzyme, a ligase, a polymerase, a fragmentase, a reverse transcriptase, or a transposase.
5. The system of claim 4 , wherein said restriction enzyme is a rare cutter.
6. The system of claim 3 , wherein said barcode comprises an oligonucleotide barcode molecule.
7. The system of claim 2 , wherein said analyte comprises a cell, a polynucleotide, a chromosome, a protein, a peptide, a polysaccharide, a sugar, a lipid, or a small molecule.
8. The system of claim 2 , wherein said analyte comprises a polynucleotide molecule.
9. The system of claim 8 , wherein said polynucleotide molecule is a deoxyribonucleic acid (DNA) molecule, a ribonucleic acid molecule, or complementary DNA (cDNA) molecule.
10. The system of claim 2 , wherein said analyte comprises a cell.
11. The system of claim 10 , wherein said partition comprises said cell and does not include any additional cells.
12. The system of claim 2 , wherein said first species is said reagent and said second species is said analyte.
13. The system of claim 2 , wherein said first species is said analyte and said second species is said reagent.
14. The system of claim 1 , wherein capsules of said plurality of capsules comprise, on average, one chromosome.
15. The system of claim 1 , wherein partitions of said plurality of partitions comprise, on average, a single capsule of said plurality of capsules.
16. The system of claim 1 , wherein said partition does not include any additional capsule of said plurality of capsules.
17. The system of claim 1 , wherein said capsule comprises an additional partition.
18. The system of claim 17 , wherein said additional partition is selected from the group consisting of an additional capsule and a droplet.
19. The system of claim 1 , wherein said capsule comprises a shell selected from the group consisting of a polymeric shell, a hydrogel, a hydrophilic shell, a hydrophobic shell, a shell with a net positive charge, a shell with a net negative charge, and a shell with a neutral charge.
20. The system of claim 1 , wherein said partition comprises a fluid that is less dense than said capsule.
21. The system of claim 1 , wherein said partition comprises a fluid that is more dense than said capsule.
22. The system of claim 1 , wherein said plurality of partitions is a plurality of aqueous droplets, wherein said first species is a cell, wherein said capsule is a gel bead, and wherein said second species is an oligonucleotide barcode molecule.
23. The system of claim 1 , wherein said capsule is configured to be degraded or disrupted by said stimulus, thereby releasing said second species into said partition.
24. The system of claim 1 , wherein said second species is an oligonucleotide barcode molecule releasably attached to said capsule.
25. The system of claim 24 , wherein said capsule comprises at least 10,000 oligonucleotide barcode molecules releasably attached thereto, wherein said at least 10,000 oligonucleotide barcode molecules comprises said oligonucleotide barcode molecule, and wherein said oligonucleotide barcode molecule is configured to release from said capsule upon application of said stimulus.
26. The system of claim 1 , wherein said capsule is a gel bead.
27. The system of claim 1 , wherein said plurality of partitions comprises a plurality of wells.
28. The system of claim 1 , wherein said change in ion concentration includes a change in pH within said capsule.
29. The system of claim 1 , wherein said partition comprises magnetic nanoparticles responsive to said magnetic stimulus.
30. The system of claim 1 , wherein said partition comprises electrically sensitive particles responsive to said electrical stimulus.
31. The system of claim 1 , wherein said partition comprises nanoparticles or chromophores responsive to said light stimulus.
32. The system of claim 1 , wherein said partition comprises photo switchable materials responsive to said light stimulus.
33. The system of claim 1 , wherein said electrical stimulus comprises application of an electric field.
34. The system of claim 1 , wherein said magnetic stimulus comprises application of a magnetic field.
35. The system of claim 1 , wherein said light stimulus comprises application of ultraviolet or visible light.
36. The system of claim 1 , wherein said stimulus is said change in ion concentration.
37. The system of claim 1 , wherein said stimulus is said reducing agent.
38. The system of claim 1 , wherein said stimulus is said light stimulus.
39. The system of claim 1 , wherein said stimulus is said electrical stimulus.
40. The system of claim 1 , wherein said stimulus is said magnetic stimulus.