Patent classifications
C12Q2545/113
Amplicon melting analysis with saturation dyes
Methods are provided for nucleic acid analysis wherein a target nucleic acid that is at least partially double stranded is mixed with a dsDNA binding dye having a percent saturation of at least 50% to form a mixture. In one embodiment, the nucleic acid is amplified in the presence of the dsDNA binding dye, and in another embodiment a melting curve is generated for the target nucleic acid by measuring fluorescence from the dsDNA binding dye as the mixture is heated. Dyes for use in nucleic acid analysis and methods for making dyes are also provided.
Nucleotide-based probes and methods for the detection and quantification of macromolecules and other analytes
Provided are unimolecular oligonucleotide probes for detecting a target in a sample. The probes use target binding-induced structural changes to detect the presence of the target in the sample. Also provided are methods of using the probes to detect a target in a sample.
Nucleotide-based probes and methods for the detection and quantification of macromolecules and other analytes
Provided are unimolecular oligonucleotide probes for detecting a target in a sample. The probes use target binding-induced structural changes to detect the presence of the target in the sample. Also provided are methods of using the probes to detect a target in a sample.
ANALYTICAL METHOD FOR QUANITIFICATION OF VIABLE BACTERIA CONTAINED IN MICROBIOTA RESTORATION THERAPY (MRT) COMPOSITIONS
Quantification of the viable bacterial microorganisms in a drug product for delivery via a gastro-nasal tube, an enema and/or a capsule or tablet. A molecular-based approach, such as PMA (propidium monazide)-qPCR (quantitative polymerase chain reaction) assays may be useful for quantification of viable bacteria. By utilizing PMA treatment in combination with qPCR, the number of viable bacterial cells in the sample can be determined.
ANALYTICAL METHOD FOR QUANITIFICATION OF VIABLE BACTERIA CONTAINED IN MICROBIOTA RESTORATION THERAPY (MRT) COMPOSITIONS
Quantification of the viable bacterial microorganisms in a drug product for delivery via a gastro-nasal tube, an enema and/or a capsule or tablet. A molecular-based approach, such as PMA (propidium monazide)-qPCR (quantitative polymerase chain reaction) assays may be useful for quantification of viable bacteria. By utilizing PMA treatment in combination with qPCR, the number of viable bacterial cells in the sample can be determined.
Diagnostic transcript and splice patterns of HPV16 in different cervical lesions
A method is described for differentiating in a subject with HPV16 between (i) a severe form of HPV16 infection and (ii) a mild form of HPV16 infection based on determining the amount of a first gene product and a second gene product in a sample of a subject and calculating a ratio of the amount of the first gene product and the amount of the second gene product. A composition is also described, including an oligonucleotide mixture, and also a kit and a device adapted to carry out the described method.
Diagnostic transcript and splice patterns of HPV16 in different cervical lesions
A method is described for differentiating in a subject with HPV16 between (i) a severe form of HPV16 infection and (ii) a mild form of HPV16 infection based on determining the amount of a first gene product and a second gene product in a sample of a subject and calculating a ratio of the amount of the first gene product and the amount of the second gene product. A composition is also described, including an oligonucleotide mixture, and also a kit and a device adapted to carry out the described method.
Method and apparatus to normalize quantitative readouts in single-cell experiments
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. One embodiment of the method includes i) selecting one or more target nucleic acid sequence of interest in an individual cell, where the target nucleic acid sequence is complementary to a nucleic acid in a cell; ii) providing a sample having a plurality of individual single cells and encapsulating one or more individual cell(s); iii) providing a sample normalization component to one or more encapsulated cell, where the normalization component comprises an exogenous nucleic acid having a known sequence; iv) providing nucleic acid primers for the target nucleic acid and the exogenous nucleic acid; v) providing a protease to each encapsulated cell and incubating the encapsulated cell with the protease in the drop to produce a cell lysate; vi) performing a nucleic acid amplification reaction to form an amplification product from the nucleic acid of a single cell, where the amplification product comprise amplicons of one or more target nucleic acid sequence and an amplicon for the exogenous nucleic acid; and vii) comparing the amplification products from the target amplicons and the exogenous nucleic acid amplicons and determining the copy number or sequence of the target nucleic acid in a single cell.
Method and apparatus to normalize quantitative readouts in single-cell experiments
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. One embodiment of the method includes i) selecting one or more target nucleic acid sequence of interest in an individual cell, where the target nucleic acid sequence is complementary to a nucleic acid in a cell; ii) providing a sample having a plurality of individual single cells and encapsulating one or more individual cell(s); iii) providing a sample normalization component to one or more encapsulated cell, where the normalization component comprises an exogenous nucleic acid having a known sequence; iv) providing nucleic acid primers for the target nucleic acid and the exogenous nucleic acid; v) providing a protease to each encapsulated cell and incubating the encapsulated cell with the protease in the drop to produce a cell lysate; vi) performing a nucleic acid amplification reaction to form an amplification product from the nucleic acid of a single cell, where the amplification product comprise amplicons of one or more target nucleic acid sequence and an amplicon for the exogenous nucleic acid; and vii) comparing the amplification products from the target amplicons and the exogenous nucleic acid amplicons and determining the copy number or sequence of the target nucleic acid in a single cell.
Method for assessing embryotoxicity
The present invention provides a method for assessing embryotoxicity of a chemical comprising: (1) a first step of measuring the expression level of one or more genes selected from among genes each comprising any of the nucleotide sequences of SEQ ID NOs: 1 to 78 and 101 to 230 and orthologous genes thereof in a sample from a non-human mammal or mammalian cell which has come into contact with a test chemical; and (2) a second step of comparing the measured value of the expression level of the gene in the sample obtained in the first step with a control value of the expression level of the gene and based on the difference assessing the level of the embryotoxicity of the test chemical in the sample; and so on.