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 method for processing a nucleic acid sample, comprising:
(a) providing a plurality of partitions, wherein a given partition of the plurality of partitions comprises the nucleic acid sample, a plurality of barcode molecules coupled to a bead, and a transposase;
(b) using the transposase to fragment the nucleic acid sample to a plurality of nucleic acid molecules in the given partition; and
(c) using barcode molecules from the plurality of barcode molecules to barcode nucleic acid molecules from the plurality of nucleic acid molecules, thereby providing barcoded nucleic acid molecules,
wherein subsequent to (a), the barcode molecules are released from the bead into the given partition.
2. The method of claim 1 , wherein the barcode molecules are released upon application of a stimulus to the given partition, the barcode molecules, or the bead.
3. The method of claim 2 , wherein the stimulus is selected from the group consisting of a change in pH, a change in ion concentration, and a reducing agent.
4. The method of claim 1 , wherein the bead includes the transposase.
5. The method of claim 1 , wherein the bead includes the nucleic acid sample.
6. The method of claim 1 , wherein the given partition includes a single cell, and wherein the nucleic acid sample is from the single cell.
7. The method of claim 1 , wherein the plurality of partitions is a plurality of droplets.
8. The method of claim 1 , wherein the plurality of partitions is a plurality of wells.
9. The method of claim 1 , wherein in (a), the nucleic acid sample is provided in the given partition in a predetermined coverage amount.
10. The method of claim 1 , further comprising, subsequent to (c), sequencing the barcoded nucleic acid molecules or derivatives thereof.
11. The method of claim 10 , wherein the sequencing comprises performing phasing on the barcoded nucleic acid molecules or derivatives thereof.
12. The method of claim 1 , further comprising removing the barcoded nucleic acid molecules or derivatives thereof from the given partition.
13. The method of claim 1 , further comprising, prior to (a):
providing a cell comprising the nucleic acid sample; and
lysing the cell, thereby generating a lysate.
14. The method of claim 13 , wherein the cell is lysed in the given partition.
15. The method of claim 13 , wherein the given partition includes a single copy of the nucleic acid sample from the cell.
16. The method of claim 1 , wherein (c) is performed in the given partition.
17. The method of claim 1 , wherein the nucleic acid sample is a chromosome or portion thereof.
18. The method of claim 1 , wherein the plurality of barcode molecules includes at least 1,000 barcode molecules comprising the same nucleic acid sequence.
19. The method of claim 1 , wherein the bead is a gel bead.
20. The method of claim 13 , wherein the lysate comprises the nucleic acid sample.
21. The method of claim 2 , wherein in at least (a), the given partition includes the stimulus.
22. The method of claim 21 , wherein the stimulus is a reducing agent.
23. The method of claim 1 , wherein each of the plurality of barcode molecules in the given partition has a sequence that is different than sequences of barcode molecules in other partitions of the plurality of partitions.
24. The method of claim 1 , wherein the plurality of barcode molecules is coupled to the bead through disulfide bonds.
25. The method of claim 10 , wherein the sequencing comprises determining linkage information in sequences generated from the barcoded nucleic acid molecules or derivatives thereof.