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US 2026/0308573 A1Published application
Nucleic Acid Sequence Analysis from Single Cells
AssigneeIllumina, Inc.
InventorsNeeraj Salathia, Jian-Bing Fan, Fiona Kaper, Gordon M. Cann, Arash Jamshidi, Alex Aravanis
Life of this patent
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Abstract
Presented herein are methods and compositions for multiplexed single cell gene expression analysis. Some methods and compositions include the use of droplets and/or beads bearing unique barcodes such as unique molecular barcodes (UMI).
Claims23
A method of preparing cDNA from a single cell comprising:
- (a) preparing a plurality of droplets, each droplet comprising (1) a reverse transcriptase, (2) a template switch oligonucleotide, and (3) one or more beads, wherein each bead comprises a plurality of oligonucleotides, each oligonucleotide comprising:
- (i) a linker;
- (ii) an amplification primer binding site;
- (iii) a bead-specific sequence that is the same on each oligonucleotide on the bead but is different on other beads, wherein said bead-specific sequence is capable of identifying the cDNA of the single cell from other cDNA;
- and
- (iv) an oligo(dT) sequence for capturing mRNAs and priming reverse transcription;
- and
- (b) performing reverse transcription and template switching within each droplet to prepare cDNA.
5 more elements
A method of preparing cDNA from a single cell comprising:
- (a) preparing a plurality of droplets, each droplet comprising (1) a reverse transcriptase, (2) a template switch oligonucleotide, and (3) one or more beads, wherein each bead comprises a plurality of oligonucleotides, each oligonucleotide comprising:
- (i) a linker;
- (ii) an amplification primer binding site;
- (iii) a unique molecular identifier (UMI);
- (iv) a bead-specific sequence that is the same on each oligonucleotide on the bead but is different on other beads, wherein said bead-specific sequence is capable of identifying the cDNA of the single cell from other cDNA;
- and
- (v) an oligo(dT) sequence for capturing mRNAs and priming reverse transcription;
- and
- (b) performing reverse transcription and template switching within each droplet to prepare cDNA.
6 more elements
Dependencies are inferred from the claim text.