Specific, accurate, and cost effective primers for performing digital PCR, compositions and kits containing the primer and methods for using and making the same are useful for detecting nucleic acid mutations. A primer useful as a first forward primer in performing digital PCR to detect a target nucleic acid in a sample, includes: a detection portion located upstream to a target sequence binding portion, and including a second forward primer binding portion having a sequence substantially complementary to a second forward primer, and a probe binding portion downstream to the second forward primer binding portion having a sequence substantially complementary to a probe; the target sequence binding portion includes a mismatch portion having a sequence not complementary to the target nucleic acid, and an amplification determinant portion downstream to the mismatch portion having a sequence complementary to a gene allele or a variant thereof encoded by the target nucleic acid.

The carborhodamine dyes disclosed herein are novel reagents suitable for automated incorporation of carborhodamine dyes into oligonucleotides that can be used in detection methods for nucleic acid targets. This disclosure provides an efficient and simple process for the preparation of carborhodamine compounds and introduces previously unknown reagents for the automated synthesis of oligonucleotide-carborhodamine conjugates.

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.

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.

Disclosed are methods, compositions and systems for detecting the presence or absence of a pathogen in a sample. The method may include the steps of obtaining a sample from the subject and treating the sample with heat to inactivate any pathogen present in the sample. The method may further include the step of treating the sample to concentrate any pathogen present in the sample. Also, the method may include isolate a pathogen-specific nucleic acid from the heat-inactivated sample and detecting the presence or absence of the isolated pathogen-specific nucleic acid. In certain embodiments, the methods and/or systems may be used to detect SARS-CoV-2. The method may employ real time RT-PCR to provide results in about 3 hours or less.

Disclosed are methods, compositions and systems for detecting the presence or absence of a pathogen in a sample. The method may include the steps of obtaining a sample from the subject and treating the sample with heat to inactivate any pathogen present in the sample. The method may further include the step of treating the sample to concentrate any pathogen present in the sample. Also, the method may include isolate a pathogen-specific nucleic acid from the heat-inactivated sample and detecting the presence or absence of the isolated pathogen-specific nucleic acid. In certain embodiments, the methods and/or systems may be used to detect SARS-CoV-2. The method may employ real time RT-PCR to provide results in about 3 hours or less.

This invention relates to methods and compositions for assessing an amount of non-native nucleic acids in a sample, such as from a subject, and/or noise, background or discordance quality check (QC). The methods and compositions provided herein can be used to determine risk of a condition, such as cancer, in a subject.

This invention relates to methods and compositions for assessing an amount of non-native nucleic acids in a sample, such as from a subject, and/or noise, background or discordance quality check (QC). The methods and compositions provided herein can be used to determine risk of a condition, such as cancer, in a subject.

The present invention relates to the detection of a target nucleic acid sequence by a PCEC (PTO Cleavage and Extension-Dependent Cleavage) assay. The present invention is characterized by generating a cleavage site for a nucleolytic enzyme on the extended duplex of which the formation is dependent on the presence of a target nucleic acid sequence. The present invention detects the occurrence of the cleavage of the extended duplex, thereby determining the presence of the target nucleic acid sequence.

The present invention relates to the detection of a target nucleic acid sequence by a PCEC (PTO Cleavage and Extension-Dependent Cleavage) assay. The present invention is characterized by generating a cleavage site for a nucleolytic enzyme on the extended duplex of which the formation is dependent on the presence of a target nucleic acid sequence. The present invention detects the occurrence of the cleavage of the extended duplex, thereby determining the presence of the target nucleic acid sequence.