Abstract: Disclosed herein are methods for analyzing one or more analytes of a cell by performing single-cell analysis. In one scenario, the one or more analytes are located on a surface of the cell. In one scenario, the one or more analytes are located internally within the cell. In one scenario, the one or more analytes include proteins located on a surface of the cell and located internally within the cell.

Abstract: The present disclosure provides methods, compositions, kits and systems for nucleic acid amplification. In some embodiments, nucleic acid amplification methods include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, nucleic acid amplification methods include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the nucleic acid amplification method employs an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel and/or in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components.

Abstract: The present invention provides methods for repurposing beads and other solid supports for separately capturing RNA and DNA without a loss of binding capacity by using a surfactant.

Abstract: Disclosed herein is a single-cell analysis workflow involving whole genome amplification for developing single-cell whole genome DNA libraries. The single-cell analysis workflow involves encapsulating and lysing cells in individual droplets and releasing genomic DNA from chromatin within the droplet. Transposases access the released genomic DNA and insert adaptor sequences into the cleaved nucleic acid fragments, thereby generating tagmented genomic DNA fragments that span the whole genome. Tagmented genomic DNA undergo nucleic acid amplification and sequencing for generating single-cell whole genome DNA libraries.

Abstract: The present invention provides methods for repurposing beads and other solid supports for separately capturing RNA and DNA without a loss of binding capacity by using a surfactant.

Abstract: The present disclosure provides methods, compositions, kits and systems for nucleic acid amplification. In some embodiments, nucleic acid amplification methods include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, nucleic acid amplification methods include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the nucleic acid amplification method employs an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel and/or in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components.

Abstract: Provided herein are methods and systems for detection of nucleic acids for single cell samples. As part of the detection, a unique step of normalization of different single cell samples is included.

Abstract: The present invention provides methods, reagents and kits for carrying out a variety of assays suitable for analyzing polynucleotides or samples that include an amplification step performed in a multiplex fashion. Also provided are methods for analyzing and improving the efficiency of amplification and for carrying out gene expression analysis.

Abstract: Digestible primers are incorporated into single cell analysis workflows to reduce and/or eliminate primer byproducts and misprimed nucleic acids. Specifically, digestible primers can participate in a first reaction, such as reverse transcription of RNA transcripts to generate cDNA, but digestible primers are digested to prevent them from participating in subsequent reactions, such as nucleic acid amplification. For example, digestible primers can include a primer with one or more ribonucleotide nucleobases, a primer with uracil bases, a primer with deoxyuridine sequences, or a primer with ribouridine sequences. Such primers can then be digested (e.g., enzymatically digested) to remove them from interfering in subsequent nucleic acid amplification reactions.

Abstract: Provided herein are methods and systems for the simultaneous targeted detection and sequencing of DNA, RNA, and Protein. In typical embodiments, the DNA, RNA, and Proteins are detected, characterized, and sequenced using just a single mammalian cell. One embodiment of detecting and characterizing DNA, RNA, or protein from a mammalian cell includes encapsulating a single cell in a drop and performing a protease digest on the encapsulated cell drop, performing a reverse transcriptase reaction; performing a droplet merger with barcoding PCR reagents and barcoding beads; performing a PCR reaction to attach the cell barcodes to the DNA targeted amplicons, RNA targeted amplicons, and protein tag amplicons, where all amplicons from the same emulsion contain the same cell barcode; and detecting and characterizing a DNA, RNA, or protein amplicon by sequencing the cell barcode incorporated into each amplicon.

Abstract: Single-cell analysis of a population of cells reveals cellular genotypes (e.g., single nucleotide variants and copy number variations) and phenotypes (e.g., protein expression) of individual cells. In one scenario, individual cells can be classified according to their respective genotypes and phenotypes. In one scenario, genotypes and phenotypes of all cells in the population are informative for identifying subpopulations of cells, thereby revealing intra-population heterogeneity. The identification of subpopulations of cells is informative for improving the understanding of cellular biology, especially in the context of diseases such as cancer, and is further informative for the better design of diagnostics and therapies.

Abstract: Single-cell analysis using combined RNA sequencing of RNA transcripts and DNA sequencing of chromatin-accessible DNA is performed to determine trajectories of single cells. Individual cells are encapsulated and lysed using reagents that do not include proteases or transposases. Cell lysates include RNA transcripts and packaged DNA (e.g., DNA packaged as chromatin) Segments of DNA in the packaged DNA are primed, amplified, and sequenced to generate sequence reads of the chromatin-accessible DNA. RNA transcripts are reverse transcribed to generate cDNA which is then primed, amplified, and sequenced to generate sequence reads. Sequence reads from the RNA-seq and DNA-seq reveal different states of cells and therefore, are useful for predicting cell trajectories.

Abstract: Provided herein are methods and systems for the simultaneous targeted detection and sequencing of DNA, RNA, and Protein. In typical embodiments, the DNA, RNA, and Proteins are detected, characterized, and sequenced using just a single mammalian cell. One embodiment of detecting and characterizing DNA, RNA, or protein from a mammalian cell includes encapsulating a single cell in a drop and performing a protease digest on the encapsulated cell drop, performing a reverse transcriptase reaction; performing a droplet merger with barcoding PCR reagents and barcoding beads; performing a PCR reaction to attach the cell barcodes to the DNA targeted amplicons, RNA targeted amplicons, and protein tag amplicons, where all amplicons from the same emulsion contain the same cell barcode; and detecting and characterizing a DNA, RNA, or protein amplicon by sequencing the cell barcode incorporated into each amplicon.

Abstract: This application relates to methods for ligating oligonucleotides having complementarity to a target nucleic acid, and amplifying the ligated oligonucleotides, where ligation and amplification occur in the same reaction mixture.

Abstract: Provided herein are methods for detection and characterization of a target nucleic acid from a single cell. One embodiment is a method for detection of a BCR-ABL gene fusion in a nucleic acid sample from a single cell having or suspected of having a BCR-ABL fusion transcript. One preferred implementation of the invention includes providing a nucleic acid amplification primer set complementary to a target nucleic acid suspected of having a BCR-ABL fusion transcript. In some embodiments, one or both primers of the nucleic acid amplification primer set have a barcode identification sequence. Also provided are methods for the detection of an AML tumor, methods are used for the detection of a leukemia, for the detection of a myeloid leukemia, and to determine the prognosis of a patient suspected of having a BCR-ABL fusion transcript.

Abstract: The present disclosure provides methods, compositions, kits and systems for nucleic acid amplification. In some embodiments, nucleic acid amplification methods include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, nucleic acid amplification methods include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the nucleic acid amplification method employs an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel and/or in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components.

Abstract: This application relates to methods for ligating oligonucleotides having complementarity to a target nucleic acid, and amplifying the ligated oligonucleotides, where ligation and amplification occur in the same reaction mixture.

Abstract: The present disclosure provides methods, compositions, kits and systems for nucleic acid amplification. In some embodiments, nucleic acid amplification methods include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, nucleic acid amplification methods include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the nucleic acid amplification method employs an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel and/or in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components.

Abstract: Provided herein are methods and systems for detection of nucleic acids for single cell samples. As part of the detection, a unique step of normalization of different single cell samples is included.

Abstract: Provided herein are methods and systems for the simultaneous targeted detection and sequencing of DNA, RNA, and Protein. In typical embodiments, the DNA, RNA, and Proteins are detected, characterized, and sequenced using just a single mammalian cell. One embodiment of detecting and characterizing DNA, RNA, or protein from a mammalian cell includes encapsulating a single cell in a drop and performing a protease digest on the encapsulated cell drop, performing a reverse transcriptase reaction; performing a droplet merger with barcoding PCR reagents and barcoding beads; performing a PCR reaction to attach the cell barcodes to the DNA targeted amplicons, RNA targeted amplicons, and protein tag amplicons, where all amplicons from the same emulsion contain the same cell barcode; and detecting and characterizing a DNA, RNA, or protein amplicon by sequencing the cell barcode incorporated into each amplicon.