Disclosed are compositions and methods for single-step PCR of a sample containing a complex target gene pool that can simultaneously amplify a wide variety of variant target gene sequences common to the sample while maintaining the original ratios of gene variants. The compositions and methods described herein utilize (1) a gene-specific primer pool that contains multiple variants that occur in a sample containing a complex mixture of target sequences that are both required for amplification of variants in the mixture which may introduce amplification bias, with (2) a non-specific PCR primer that is designed to target multiple gene-specific primer variants and eliminate amplification bias.

Methods of detecting and quantifying target molecules, such as proteins, in a biological sample are provided. The disclosed methods include capturing target molecules with aptamers, replacing the aptamers with aptamer identification sequences, and then sequencing the aptamer identification sequences using next-generation sequencing techniques.

Methods of detecting and quantifying target molecules, such as proteins, in a biological sample are provided. The disclosed methods include capturing target molecules with aptamers, replacing the aptamers with aptamer identification sequences, and then sequencing the aptamer identification sequences using next-generation sequencing techniques.

Disclosed herein is a method of quantifying a mutant allele burden of a target gene in a subject. The method includes providing a first plasmid that includes a mutant allele sequence and an internal control sequence, and a second plasmid that includes a wild-type allele sequence and the internal control sequence, and subjecting DNA of the subject to quantitative polymerase chain reaction to measure a mutant allele expression level of the target gene, so as to determine the mutant allele burden of the target gene in the subject based on a standard curve of the mutant allele burden of the target gene created by serial dilution of the first and second plasmids.

Disclosed herein is a method of quantifying a mutant allele burden of a target gene in a subject. The method includes providing a first plasmid that includes a mutant allele sequence and an internal control sequence, and a second plasmid that includes a wild-type allele sequence and the internal control sequence, and subjecting DNA of the subject to quantitative polymerase chain reaction to measure a mutant allele expression level of the target gene, so as to determine the mutant allele burden of the target gene in the subject based on a standard curve of the mutant allele burden of the target gene created by serial dilution of the first and second plasmids.

Methods and systems for mRNA analysis and quantification of mRNA expression in cells are provided. An example method includes introducing a first capture probe and a second capture probe into the cells, the first capture probe and the second capture probe each configured to be complementary to a respective section of target mRNA within the cells, wherein binding of the first and second capture probes to the respective sections of the target mRNA results in tagging of the cells and causes the first and second capture probes to form clusters with each other. The first capture probe and the second capture probe are each bound to magnetic nanoparticles (MNPs) that, when trapped within the tagged cells, cause the tagged cells to be susceptible to magnetic forces. The method and system further include introducing the cells into a device configured to magnetically capture tagged cells.

Methods and systems for mRNA analysis and quantification of mRNA expression in cells are provided. An example method includes introducing a first capture probe and a second capture probe into the cells, the first capture probe and the second capture probe each configured to be complementary to a respective section of target mRNA within the cells, wherein binding of the first and second capture probes to the respective sections of the target mRNA results in tagging of the cells and causes the first and second capture probes to form clusters with each other. The first capture probe and the second capture probe are each bound to magnetic nanoparticles (MNPs) that, when trapped within the tagged cells, cause the tagged cells to be susceptible to magnetic forces. The method and system further include introducing the cells into a device configured to magnetically capture tagged cells.

The present invention provides a method for quantitatively detecting virus-derived RNA and/or DNA in an environmental sample or a fecal sample with high sensitivity, and a kit for quantitatively detecting virus-derived RNA and/or DNA in an environmental sample or a fecal sample with high sensitivity.

The present invention provides a method for quantitatively detecting virus-derived RNA and/or DNA in an environmental sample or a fecal sample with high sensitivity, and a kit for quantitatively detecting virus-derived RNA and/or DNA in an environmental sample or a fecal sample with high sensitivity.

A method is provided comprising the following steps: (a) treating a nucleic acid with one or more bisulphites to convert non-methylated cytosines in the nucleic acid into uracils while leaving methylated cytosines unchanged to form a treated nucleic acid strand that is part of two joined nucleic acid strands; (b) ligating a first adapter to a 3 end of the treated nucleic acid strand to thereby form a once adapter ligated nucleic acid strand, the first adapter having a first protruding random sequence that is at least 3 bases long and that acts as a splint for the two joined nucleic acid strands; (c) ligating a second adapter to a 5 end of the once adapter ligated nucleic acid strand to thereby form a twice ligated nucleic acid strand, the second adapter having a second protruding random sequence that is at least 3 bases long and that acts as a splint for the two joined nucleic acid strands; and (d) performing polymerase chain reaction (PCR) amplification on the twice ligated nucleic acid strand to thereby generate copies of the twice ligated nucleic acid strand.