The subject invention pertains to the detection and differentiation of genetic variations by nucleic acid amplification. The invention provides methods of detecting one or more genetic variations in a nucleic acid that are in close proximity simultaneously. The invention further provides primer and probe oligonucleotides and methods of using said primers and probes in assays to detect genetic variants of concern of SARS-CoV-2. The methods of the invention detect genetic variants of other pathogens, including influenza, or genetic variants involved in inheritable diseases or cancer.

The subject invention pertains to the detection and differentiation of genetic variations by nucleic acid amplification. The invention provides methods of detecting one or more genetic variations in a nucleic acid that are in close proximity simultaneously. The invention further provides primer and probe oligonucleotides and methods of using said primers and probes in assays to detect genetic variants of concern of SARS-CoV-2. The methods of the invention detect genetic variants of other pathogens, including influenza, or genetic variants involved in inheritable diseases or cancer.

Disclosed are methods for performing capillary electrophoresis on two or more nucleic acid samples. The methods employ a forward voltage to move a first sample forward from an inlet to an interrogation region in the capillary, then a backward voltage to move the first sample backward, and then a forward voltage again to move the first sample and a second sample forward. Systems and apparatuses for performing capillary electrophoresis are also provided.

Disclosed are methods for performing capillary electrophoresis on two or more nucleic acid samples. The methods employ a forward voltage to move a first sample forward from an inlet to an interrogation region in the capillary, then a backward voltage to move the first sample backward, and then a forward voltage again to move the first sample and a second sample forward. Systems and apparatuses for performing capillary electrophoresis are also provided.

The present invention provides a new and improved multiplexed oligonucleotide detection method based on the nanopore technology with one or more probes containing a sequence with complementarity to the target oligonucleotide, a terminal extension at the probe's 3′ terminus, 5′ terminus, or both termini and a label attached to the terminus. The improved probes and probe sets enable sensitive, selective, and direct multiplex detection, differentiation and quantification of distinct target oligonucleotides such as miRNAs. The inventive detection method may also be employed as a non-invasive and cost-effective diagnostic method based on miRNA levels in the patient's tissue sample.

The present invention provides a new and improved multiplexed oligonucleotide detection method based on the nanopore technology with one or more probes containing a sequence with complementarity to the target oligonucleotide, a terminal extension at the probe's 3′ terminus, 5′ terminus, or both termini and a label attached to the terminus. The improved probes and probe sets enable sensitive, selective, and direct multiplex detection, differentiation and quantification of distinct target oligonucleotides such as miRNAs. The inventive detection method may also be employed as a non-invasive and cost-effective diagnostic method based on miRNA levels in the patient's tissue sample.

The present disclosure provides methods and kits for performing high multiplex PCR using molecular barcodes. The methods disclosed herein separately extend a set of primers (BC primers) that each comprise a target-specific sequence, a molecular barcode and a universal sequence, and amplify the resulting extension products using another set of primers (LA primers) that each comprise another target-specific sequence and a universal sequence. The methods may further comprise amplification using universal primers (preferably comprising an adapter).

The present disclosure provides methods and kits for performing high multiplex PCR using molecular barcodes. The methods disclosed herein separately extend a set of primers (BC primers) that each comprise a target-specific sequence, a molecular barcode and a universal sequence, and amplify the resulting extension products using another set of primers (LA primers) that each comprise another target-specific sequence and a universal sequence. The methods may further comprise amplification using universal primers (preferably comprising an adapter).

The present invention is concerned with PCR-based detection methods and kits for the identification, differentiation, and quantification of different bacterial strains (e.g., Gram-negative bacterial strains), and also association of two or more PCR-positive genes to a single genome. The methods generally comprise carrying out PCR reactions using at least a first PCR primer set and/or probe for at least one target nucleic acid; and a second PCR primer set and/or probe for at least a second target nucleic acid. Positive PCR reaction products are then detected to determine test samples containing positive PCR reaction products for both the first and second target nucleic acids. This information can be used to calculate the gene association rate to determine whether the sample contains, for example, Shiga toxin-producing E. coli of the O-type serogroup.

The present disclosure relates to methods and compositions involving HCR reactions that involve initiators that are split into two or more parts. Effective HCR is dependent upon two or more of these split initiators being brought into proximity (e.g., via binding events mediated by a target) such that a full initiator is formed that is capable of triggering HCR signal amplification.