A thermal cycling device comprising a number of fixed thermal zones and a fixed conduit passing through the thermal zones. A controller maintains each thermal zone including its section of conduit at a constant temperature. A series of droplets flows through the conduit so that each droplet is thermally cycled, and a detection system detects fluorescence from droplets at all of the thermal cycles. The conduit is in a single plane, and so a number of thermal cycling devices may be arranged together to achieve parallelism. The flow conduit comprises a channel and a capillary tube inserted into the channel. The detection system may perform scans along a direction to detect radiation from a plurality of cycles in a pass.

Disclosed herein is an apparatus comprising: a probe carrier, an optical system and a sensor; wherein the probe carrier comprises a substrate, a first layer and a second layer; wherein the substrate comprises a first surface, a second surface, one or more locations on the first surface configured to be deposit sites for one or more probes; wherein the second surface is at an opposite side of the substrate from the first surface; wherein the first layer is on the first surface of the substrate or is embedded in the substrate under the first surface; wherein the second layer is on the second surface of the substrate or is embedded in the substrate under the second surface; the first and second layers are configured to reduce crosstalk between probes at different locations.

Enhanced composite liquid cell (CLC) devices and methods of using the same are provided. The devices find use in, among other applications, CLC mediated nucleic acid library generation protocols, e.g., for use in next generation sequencing applications.

The present invention relates to a method and system for Polymerase Chain Reaction (“PCR”), High Resolution Melt (“HRM”) analysis and microfluidics, and, more specifically, to a method and system for implementing the processes of PCR and HRM on a microscale in a microfluidics chamber for certain purposes including for purposes of DNA detection and/or extraction.

The present disclosure is drawn to temperature-cycling microfluidic devices. In one example, a temperature-cycling microfluidic device can include a driver chip having a top surface and a heat exchange substrate having a top surface coplanar with the top surface of the driver chip. A fluid chamber can be located on the top surface of the driver chip. A first and second microfluidic loop can have fluid driving ends and fluid outlet ends connected to the fluid chamber and can include portions thereof located on the top surface of the heat exchange substrate. A first and second fluid actuator can be on the driver chip. The first and second fluid actuators can be associated with the fluid driving ends of the first and second microfluidic loops, respectively, to circulate fluid through the first and second microfluidic loops.

A nucleic acid amplification reaction method includes subjecting a reaction mixture containing a nucleic acid amplification reaction reagent to be used for amplifying a nucleic acid to a thermal cycle for amplifying the nucleic acid, wherein in the thermal cycle, a heating time for an annealing reaction and an elongation reaction is 1 sec or more and 10 sec or less, the nucleic acid amplification reaction reagent contains a forward primer, a reverse primer, a polymerase, and a fluorescently labeled probe, the concentration of the forward primer is 0.4 μM or more and 3.2 μM or less, the concentration of the reverse primer is 0.4 μM or more and 3.2 μM or less, the amount of the polymerase is 0.5 U or more and 4 U or less, and the concentration of the fluorescently labeled probe is 0.15 μM or more and 1.2 μM or less.

Aspects of the present disclosure include sample preparation cartridges including a frame that includes a plurality of wells integrated therewith, where the plurality of wells have a closed bottom and an open top. The frame further includes an opening within the frame having a reaction vessel (RV) or RV cap removably disposed therein, where the plurality of wells and the opening are linearly arranged relative to each other. Also provided are sample preparation cartridges that include a frame, two or more cartridge separation projections on a top side of the frame, and two or more cartridge separation projections on a bottom side of the frame. The cartridge separation projections separate the cartridge and a different cartridge when the cartridge and different cartridge are stacked. Methods of using the sample preparation cartridges, as well as nucleic acid sample preparation units that include the sample preparation cartridges, are also provided.

The present invention provides a nucleic acid sequence amplification method and apparatuses thereof that are simple in the design and easy to miniaturize and integrate into complex apparatuses, with capability of using DNA polymerases that are not thermostable. In the present invention, a plurality of heat sources are combined to supply or remove heat from specific regions of the sample such that a specific spatial temperature distribution is maintained inside the sample by locating a relatively high temperature region lower in height than a relatively low temperature region.

A biological sample analysis system including a sample preparation system forming droplets of segmented sample separated by a carrier fluid immiscible with the sample. The droplets include reaction mixtures for amplification of at least one target nucleic acid. A thermal cycling device having a sample block having a plurality of controlled thermal zones, and a containment structure in thermal communication with the plurality of controlled thermal zones. The containment structure receives and contains the droplets of segmented sample separated by the immiscible carrier fluid from the sample preparation system. A controller for controlling a temperature in each thermal zone of the sample block. A detection system detects electromagnetic radiation emitted from each of the droplets individually from the queue of droplets as they flow past the detection system. A positioning system to facilitate moving a queue of the droplets in the thermal cycling device relative to the detection system.

Methods and containers are provided for identifying a species, illustratively a bacterial species. Illustrative methods comprise amplifying various genes in the nucleic acid from the bacterial species in a single reaction mixture using pairs of outer first-stage primers designed to hybridize to generally conserved regions of the respective genes to generate a plurality of first-stage amplicons, dividing the reaction mixture into a plurality of second-stage reactions, each using a unique pair of second-stage primers, each pair of second-stage primers specific for a target bacterial species or subset of bacterial species, detecting which of the second-stage reactions amplified, and identifying the bacterial species based on second-stage amplification. Methods for determining antibiotic resistance are also provided, such methods also using first-stage primers for amplifying genes known to affect antibiotic resistance a plurality of the second-stage reactions wherein each pair of second-stage primers specific for a specific gene for conferring antibiotic resistance.