Methods for detection and quantification of nucleic acid or protein targets in a sample
AssigneeFluidigm Corporation
InventorsMichael Lucero, Marc Unger
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Abstract
The invention provides an assay method for detection and/or quantification of a plurality of nucleic acid or protein targets in a sample. In the method probes are used to associate a detectable tag sequence with each of the selected targets present in the sample. Probes or primers sufficient to identify at least 25, and preferably at least 500, different targets are used. The method involves segregating aliquots of the sample from each other and detecting the tag sequences in each aliquot.
Claims18
An assay method for detection of a plurality of nucleic acid or protein targets in a sample comprising
- combining target-specific probes with the sample under conditions in which the probe binds a target, wherein target-specific probes sufficient to identify at least 25 different targets are used, wherein each probe comprises a nucleic acid tag sequence, and wherein the target-specific probe forms a target-specific probe product associated with the target if the target is present in the sample, thereby associating the tag with the target;
- distributing aliquots of the sample to separate compartments of a microfluidic device and segregating the aliquots from each other;
- querying each aliquot of the sample for the presence of a different target-specific probe product, wherein said querying comprises detecting the nucleic acid tag sequence if present in the aliquot.
An assay method for detection of a plurality of DNA targets in a sample comprising
- combining target-specific padlock probes sufficient to identify at least 25 different targets with the sample under conditions in which the probes bind and cantenate to the target molecules, if present, forming a target-specific probe product
- wherein each probe comprises a nucleic acid tag sequence and wherein the padlock probes form target-specific probe products associated with the target if the target is present in the sample, thereby associating the tag sequence with the target
- distributing aliquots of the sample to separate compartments of a microfluidic device and segregating the aliquots from each other;
- querying each aliquot for the presence of a different probe molecule catenated to the target, wherein said querying comprises detecting the tag sequence if present in the aliquot, and wherein detecting the tag comprises amplifying the tag sequence using PCR primers and detecting the amplification using a dual-labeled fluorogenic oligonucleotide probe.
Dependencies are inferred from the claim text.