Methods for nucleic acid analysis
AssigneeUltima Genomics, Inc.
InventorsGilad Almogy, Florian Oberstrass, Omer Barad, Chandan Shee
Life of this patent
Estimating the term…
Abstract
The present disclosure provides methods and processes for increasing the efficiency and accuracy of nucleic acid sequencing using techniques such as polymerase chain reaction (PCR). The methods described herein can be used to achieve clonal amplification even with a greater than Poisson distribution of beads and/or nucleic acid templates into an emulsion. A PCR method may comprise generating a partition (e.g., a droplet) comprising at least two beads and/or at least two nucleic acid molecules and generating clonal amplification products corresponding to the nucleic acid molecule, at least a subset of which may be attached to a bead.
Claims19
A method for nucleic acid processing, comprising:
- (a) providing a plurality of partitions, wherein a partition of said plurality of partitions comprises (i) at least two beads of a plurality of beads, comprising a first bead and a second bead, wherein said first bead comprises a first primer and said second bead comprises a second primer, wherein said first primer and said second primer have different sequences, (ii) at least two nucleic acid molecules of a plurality of nucleic acid molecules, comprising a first nucleic acid molecule and a second nucleic acid molecule, wherein said first nucleic acid molecule and said second nucleic acid molecule have different nucleic acid sequences, wherein said first nucleic acid molecule comprises a first adaptor and said second nucleic acid molecule comprises a second adaptor, wherein said first adaptor and said second adaptor have different sequences, wherein said first primer has sequence complementarity with said first adaptor and said second primer has sequence complementarity with said second adaptor, and (iii) one or more reagents;
- (b) in said partition, with said first primer hybridized to said first adaptor and said second primer hybridized to said second adaptor, (i) using said first nucleic acid molecule and said one or more reagents to generate one or more first amplification products of said first nucleic acid molecule, wherein at least a subset of said one or more first amplification products is attached to said first bead, and (ii) using said second nucleic acid molecule and said one or more reagents to generate one or more second amplification products of said second nucleic acid molecule, wherein at least a subset of said one or more second amplification products is attached to said second bead;
- (c) recovering said first bead and said second bead from said partition;
- and
- (d) assaying (i) a first amplification product of said at least said subset of said one or more first amplification products attached to said first bead, to identify a sequence of said first nucleic acid molecule and (ii) a second amplification product of said at least said subset of said one or more second amplification products attached to said second bead, to identify a sequence of said second nucleic acid molecule.
Dependencies are inferred from the claim text.