Systems, methods, and apparatuses are provided for self-contained nucleic acid preparation, amplification, and analysis.

A reaction processor is provided with a reaction processing vessel in which a channel is formed, a liquid feeding system, a temperature control system for providing a high temperature region and a low temperature region to the channel, and a fluorescence detector for detecting the sample passing through a fluorescence detection region of the channel, and a CPU for controlling the liquid feeding system based on a signal that is detected. A target stop position X.sup.[L].sub.0(n+1) of the sample in the low temperature region in an (n+1)th cycle is corrected from a target stop position X.sup.[L].sub.0(n) of the sample in the low temperature region in the nth cycle based on the result of stopping control on the sample in the nth cycle.

An apparatus includes a substrate, a first heating element, and a second heating element. The substrate includes a first portion, a second portion, and a third portion that is between the first portion and the second portion. The first portion is characterized by a first thermal conductivity, the second portion is characterized by a second thermal conductivity, and the third portion is characterized by a third thermal conductivity. The third thermal conductivity is less than the first thermal conductivity and the second thermal conductivity. The first heating element is coupled to the first portion of the substrate, and is configured to produce a first thermal output. The second heating element is coupled to the second portion of the substrate, and configured to produce a second thermal output. The second thermal output is different from the first thermal output.

A reaction processor includes: a reaction processing vessel including a channel in which a sample moves and a pair of air communication ports, a first air communication port and a second air communication port, provided at respective ends of the channel; a temperature control system that provides a medium temperature region and a high temperature region between the first air communication port and the second air communication port in the channel; and a liquid feeding system that discharges and sucks air in order to move and stop the sample inside the channel. One of the pair of air communication ports of the reaction processing vessel that is farther away from the high temperature region communicates with the liquid feeding system via a tube. One of the pair of air communication ports of the reaction processing vessel that is closer to the high temperature region is opened to atmospheric pressure.

Methods are provided for separating magnetically responsive beads from a droplet in a droplet actuator. Droplet operations electrodes and a magnet are arranged in a droplet actuator to manipulate a bead-containing droplet and position it relative to a magnetic field region that attracts the magnetically responsive beads. The droplet operations electrodes are operated to control the droplet shape and transport it away from the magnetic field region to form a concentration of beads in the droplet. The continued transport of the droplet away from the magnetic field causes the concentration of beads to break away from the droplet to yield a small, concentrated bead-containing droplet immobilized by the magnet.

Provided is a digital microfluidic device for quick polymerase chain reaction. The digital microfluidic device includes an enclosed chamber for holding droplets comprising PCR mixtures. The chamber has an upper layer and a lower layer, which provide a top heater and a bottom heater contained in a thermal electrode respectively to form dual heaters. The lower layer further has an array of electrodes and a dielectric layer, e.g. Norland Optical adhesive 61, coating thereon. Such arrangement of the digital microfluidic device allows quick and homogeneous heating of droplets to lower the heating voltage, shorten the reaction time, and prevent the dielectric layer from breakdown during the thermal cycle.

The present invention relates to systems and methods for the arrangement of droplets in pre-determined locations. Many applications require the collection of time-resolved data. Examples include the screening of cells based on their growth characteristics or the observation of enzymatic reactions. The present invention provides a tool and related techniques which addresses this need, and which can be used in many other situations. The invention provides, in one aspect, a tool that allows for stable storage and indexing of individual droplets. The invention can interface not only with microfluidic/microscale equipment, but with macroscopic equipment to allow for the easy injection of liquids and extraction of sample droplets, etc.

Instrument for performing a diagnostic test on a fluidic cartridge A cartridge reader is for carrying out a diagnostic test on a sample contained in a fluidic cartridge inserted into the reader. The fluidic cartridge comprises a fluidic layer comprising at least one sample processing region, at least one collapsible blister containing a liquid reagent, a pneumatic interface, an electrical interface and at least one mechanical valve. The reader comprises a housing; an upper clamp occupying a fixed position relative to the reader, and a lower clamp, movable relative to the first clamp, wherein the upper clamp and the lower clamp define a cartridge receiving region therebetween. The reader comprises a thermal module comprised in the lower clamp, wherein the thermal module comprises at least one thermal stack for heating the at least one sample processing region of the cartridge inserted into the reader. The reader comprises at least one mechanical actuator for actuating the mechanical valve comprised in the cartridge inserted into the reader.

Microchannels include membranes operable with magnets or other actuators to deliver samples to one or more reaction zones defined in the microchannels. Membrane flexing can direct samples to selected reaction zones and each reaction zone can be independently temperature controlled to implement a PCR-based sample analysis.

Method of analysis. In the method, a first emulsion and a second emulsion substantially separated from one another by a spacer fluid may be formed. The first emulsion, the spacer fluid, and the second emulsion may be flowed in a channel from a fluid inlet to a fluid outlet of a heating and cooling station having two or more temperature-controlled zones, such that each emulsion is thermally cycled to promote amplification of a nucleic acid target in droplets of the emulsion. Amplification data may be collected from individual droplets of each emulsion downstream of the heating and cooling station. A level of the nucleic acid target present in each emulsion may be determined based on the amplification data collected from the individual droplets of the emulsion.