Provided are systems and methods of sample preparation and analyte detection.

Provided herein are engineered variants of archaeal, prokaryotic, and eukaryotic polymerases that exhibit enhanced thermostability, enhanced incorporation of 3′ modified nucleotides, and improved uracil-tolerance, in polymerase-catalyzed nucleotide extension reactions relative to wild type polymerase enzymes. Also provided are uses of the engineered polymerases for forming complexed polymerases, forming binding complexes and forming ternary complexes, and uses for conducting nucleic acid sequencing reactions.

Provided herein are engineered variants of archaeal, prokaryotic, and eukaryotic polymerases that exhibit enhanced thermostability, enhanced incorporation of 3′ modified nucleotides, and improved uracil-tolerance, in polymerase-catalyzed nucleotide extension reactions relative to wild type polymerase enzymes. Also provided are uses of the engineered polymerases for forming complexed polymerases, forming binding complexes and forming ternary complexes, and uses for conducting nucleic acid sequencing reactions.

The invention features methods panels, cartridges, and systems for detecting pathogens and for diagnosing and treating diseases, including bacteremia and sepsis.

The invention features methods panels, cartridges, and systems for detecting pathogens and for diagnosing and treating diseases, including bacteremia and sepsis.

There is provided a paper-based colorimetric sensor kit for quickly and simply diagnosing mercury in situ with a naked eye. The paper-based colorimetric sensor kit includes: a circular template for rolling circle amplification (RCA); a primer that does not hybridize to the circular template when a mercury ion is bonded to a primer that hybridizes to the circular template; a DNA polymerase; a sensing material kit including a nanoparticle probe labeled to a DNA coil formed in the circular template for RCA; and a radial chromatography paper.

There is provided a paper-based colorimetric sensor kit for quickly and simply diagnosing mercury in situ with a naked eye. The paper-based colorimetric sensor kit includes: a circular template for rolling circle amplification (RCA); a primer that does not hybridize to the circular template when a mercury ion is bonded to a primer that hybridizes to the circular template; a DNA polymerase; a sensing material kit including a nanoparticle probe labeled to a DNA coil formed in the circular template for RCA; and a radial chromatography paper.

Provided herein are compositions and methods for identifying or quantitating one or more analytes in sample. The composition can comprise an affinity molecule reversibly conjugated to a label moiety via a double-stranded nucleic acid linker or via an adaptor molecule. The affinity molecule and the label moiety can be linked to different strands of the double-stranded nucleic acid linker. Compositions can be used in any biological assays for detection, identification and/or quantification of target molecules or analytes, including multiplex staining for molecular profiling of individual cells or cellular populations. For example, the compositions can be adapted for use in immunofluorescence, fluorescence in situ hybridization, immunohistochemistry, western blot, and the like.

Provided herein are compositions and methods for identifying or quantitating one or more analytes in sample. The composition can comprise an affinity molecule reversibly conjugated to a label moiety via a double-stranded nucleic acid linker or via an adaptor molecule. The affinity molecule and the label moiety can be linked to different strands of the double-stranded nucleic acid linker. Compositions can be used in any biological assays for detection, identification and/or quantification of target molecules or analytes, including multiplex staining for molecular profiling of individual cells or cellular populations. For example, the compositions can be adapted for use in immunofluorescence, fluorescence in situ hybridization, immunohistochemistry, western blot, and the like.

A digital assay for a micro RNA (miRNA) or other target analyte in a sample makes use of nanoparticles that absorb light at the resonant wavelength of a photonic crystal (PC). Such nanoparticles locally quench the resonant reflection of light from the PC when present on the surface of the PC. The nanoparticles are functionalized to specifically bind to the target analyte, and the PC surface is functionalized to specifically bind to the nanoparticles that have bound to the target analyte. The sample is exposed to the functionalized nanoparticles, and the individual nanoparticles bound to the PC surface can be identified and counted based on reduced intensity values in the reflected light from the PC. The number of bound nanoparticles that are counted in this way can be correlated to the abundance of the target analyte in the sample.