Provided are a sequencing library and applications thereof. The provided sequencing library includes a first nucleic acid molecule and a second nucleic acid molecule. The first nucleic acid molecule carries a cell index sequence and a droplet index sequence. The second nucleic acid molecule carries an insert fragment and a cell index sequence.

Provided are a sequencing library and applications thereof. The provided sequencing library includes a first nucleic acid molecule and a second nucleic acid molecule. The first nucleic acid molecule carries a cell index sequence and a droplet index sequence. The second nucleic acid molecule carries an insert fragment and a cell index sequence.

Provided are devices, systems, and methods for the identification, quantification, and profiling of microscopic organisms. The methods for the identification, quantification, and profiling of microscopic organisms include, for example, the selective enrichment of microscopic organisms from a heterogeneous sample; subsequent loading of the microscopic organisms into microfluidic channels or reaction chambers; direct amplification of nucleic acids from single, isolated microscopic organisms; and examination of amplification products using digital High Resolution Melting (HRM) analysis.

Provided are devices, systems, and methods for the identification, quantification, and profiling of microscopic organisms. The methods for the identification, quantification, and profiling of microscopic organisms include, for example, the selective enrichment of microscopic organisms from a heterogeneous sample; subsequent loading of the microscopic organisms into microfluidic channels or reaction chambers; direct amplification of nucleic acids from single, isolated microscopic organisms; and examination of amplification products using digital High Resolution Melting (HRM) analysis.

An embodiment of a system includes a compartment-generating device, a compartment detector, and electronic computing circuitry. The device is configured to generate compartments of a digital assay, at least one of the compartments having a respective volume that is different from a respective volume of each of at least another one of the compartments. The detector is configured to determine a number of the compartments each having a respective number of a target that is greater than a threshold number of the target. And the electronic circuitry is configured to determine a bulk concentration of the target in a source of the sample in response to the determined number of compartments. Because such a system can be configured to estimate a bulk concentration of a target in a source from a polydisperse digital assay, the system can be portable, and lower-cost and faster, than conventional systems.

An embodiment of a system includes a compartment-generating device, a compartment detector, and electronic computing circuitry. The device is configured to generate compartments of a digital assay, at least one of the compartments having a respective volume that is different from a respective volume of each of at least another one of the compartments. The detector is configured to determine a number of the compartments each having a respective number of a target that is greater than a threshold number of the target. And the electronic circuitry is configured to determine a bulk concentration of the target in a source of the sample in response to the determined number of compartments. Because such a system can be configured to estimate a bulk concentration of a target in a source from a polydisperse digital assay, the system can be portable, and lower-cost and faster, than conventional systems.

An example system includes an array of retaining features in a microfluidic cavity, an array of thermally controlled releasing features, and a controller coupled to each releasing feature in the array of releasing feature. Each retaining feature in the array of retaining features is to position capsules at a predetermined location, the capsules having a thermally degradable shell enclosing a biological reagent therein. Each releasing feature in the array of releasing features corresponds to a retaining feature and is to selectively cause degradation of the shell of a capsule. Each releasing feature is to generate thermal energy to facilitate degradation of the shell. The controller is to selectively activate at least one releasing feature in the array of thermally controlled releasing features to release the biological reagent in the capsules positioned at the retaining feature corresponding to the activated releasing feature.

An example system includes an array of retaining features in a microfluidic cavity, an array of thermally controlled releasing features, and a controller coupled to each releasing feature in the array of releasing feature. Each retaining feature in the array of retaining features is to position capsules at a predetermined location, the capsules having a thermally degradable shell enclosing a biological reagent therein. Each releasing feature in the array of releasing features corresponds to a retaining feature and is to selectively cause degradation of the shell of a capsule. Each releasing feature is to generate thermal energy to facilitate degradation of the shell. The controller is to selectively activate at least one releasing feature in the array of thermally controlled releasing features to release the biological reagent in the capsules positioned at the retaining feature corresponding to the activated releasing feature.

A method of forming a polymer matrix array includes treating a surface within a well of a well array with a surface compound including a surface reactive functional group and a radical-forming distal group; applying an aqueous solution including polymer precursors to the well of the well array; and activating the radical-forming distal group of the surface coupling compound with an initiator and atom transfer radical polymerization (ATRP) catalyst to initiate radical polymerization of the polymer precursors within the well of the well array to form the polymer matrix array.

A method of forming a polymer matrix array includes treating a surface within a well of a well array with a surface compound including a surface reactive functional group and a radical-forming distal group; applying an aqueous solution including polymer precursors to the well of the well array; and activating the radical-forming distal group of the surface coupling compound with an initiator and atom transfer radical polymerization (ATRP) catalyst to initiate radical polymerization of the polymer precursors within the well of the well array to form the polymer matrix array.