METHODS AND SYSTEMS FOR PROCESSING POLYNUCLEOTIDES
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 nucleic acid preparation or analysis, comprising:
(a) partitioning a plurality of barcode molecules and a plurality of nucleic acid molecules derived from a cell-free biological sample in a plurality of partitions, wherein said plurality of barcode molecules comprise a plurality of barcode sequences;
(b) in said plurality of partitions, using said plurality of nucleic acid molecules and said plurality of barcode molecules to generate a plurality of barcoded nucleic acid molecules, which plurality of barcoded nucleic acid molecules comprises said plurality of barcode sequences;
(c) recovering or releasing said plurality of barcoded nucleic acid molecules from said plurality of partitions.
2 . The method of claim 1 , wherein a barcode sequence associated with a given partition of said plurality of partitions is different from a barcode sequence associated with other partitions of said plurality of partitions.
3 . The method of claim 1 , wherein said plurality of partitions is a plurality of droplets.
4 . The method of claim 1 , wherein said plurality of partitions is a plurality of wells.
5 . The method of claim 1 , wherein said plurality of barcode molecules are coupled to beads.
6 . The method of claim 5 , wherein said beads are solid particles.
7 . The method of claim 5 , wherein said beads are gel beads.
8 . The method of claim 5 , further comprising releasing said plurality of barcode molecules from said beads in said plurality of partitions.
9 . The method of claim 5 , wherein said plurality of barcode molecules is covalently attached to said beads through linkers.
10 . The method of claim 1 , wherein said plurality of nucleic acid molecules are ribonucleic acid molecules.
11 . The method of claim 1 , wherein said plurality of nucleic acid molecules are deoxyribonucleic acid molecules.
12 . The method of claim 1 , wherein said cell-free biological sample is a bodily fluid sample.
13 . The method of claim 12 , wherein said bodily fluid sample is blood.
14 . The method of claim 12 , wherein said bodily fluid sample is serum.
15 . The method of claim 12 , wherein said bodily fluid sample is saliva.
16 . The method of claim 12 , wherein said bodily fluid sample is urine.
17 . The method of claim 1 , wherein in (c), said plurality of barcoded nucleic acid molecules is generated by nucleic acid amplification using said plurality of nucleic acid molecules and said plurality of barcode molecules.
18 . The method of claim 1 , wherein a partition of said plurality of partitions comprises barcode molecules from said plurality of barcode molecules, and wherein said barcode molecules comprise a common barcode sequence.
19 . The method of claim 1 , further comprising, subsequent to (c), sequencing said plurality of barcoded nucleic acid molecules or derivatives thereof to generate a plurality of sequences, which plurality of sequences comprise sequences of said plurality of nucleic acid molecules derived from said cell-free biological sample and said plurality of barcode sequences.
20 . The method of claim 19 , further comprising using said plurality of sequences to differentiate between haplotypes in said plurality of nucleic acid molecules derived from said cell-free biological sample.