The present invention is based on the discovery of sensitive and specific methylation detection by a) controlling excessive DNA degradation prior to conversion by incubating DNA conversion reagent (e.g. bisulfite reagent) directly with a nucleic acid containing sample without requiring prior nucleic acid purification from the sample and without requiring prior nucleic acid denaturation at elevated temperatures of 98° C. and, b) optimizing bisulfite removal by controlling the pumping rate flow of the bisulfite treated sample over an extraction membrane inside an automated system. In some embodiments, a cartridge for the detection of methylated DNA is provided.

Described herein are devices, systems, fluidic devices, kits, and methods for detection of target nucleic acids.

Thermal cycling apparatus for polymerase chain reaction (PCR) of nucleic acid is provided. Bath media in a first bath and a second bath are maintainable at two different temperatures. A transfer means allows the reactor to be in the two baths in a plurality of thermal cycles to alternately attain a predetermined high target temperature T.sub.HT and a predetermined low target temperature T.sub.LT. A florescent imaging means images the reaction material during the thermal cycling. A powder-cleaning device mechanically removes particles of powder that adhere to the reactor, when powder is the bath medium in use in at least one of the baths.

An apparatus, multi-well plate and method for automated cell lysis and nucleic acid purification and amplification. The plate includes a lysis well, at least one wash well, an elution well, and a PCR chip. The apparatus includes a vertically aligned rotor mixer comprising a magnetic tip and actuators for moving the rotor mixer in a vertical and horizontal directions, to transfer magnetic beads from well to well. The rotor mixer is used to vortex lysis mixtures, wherein the vortexing speed is sufficient to overcome the magnetic attraction between the beads and mixer tip and disperse the beads in solution, to collect nucleic acids such as DNA in an elution solution that is transferred to the PCR chip for amplification of target sequences.

Examples herein involve sorting a droplet including a biologic sample. In a particular example, sorting a droplet including a biologic sample includes generating a droplet including a biologic sample and a pH sensitive surfactant, and heating a nucleic acid molecule in the biologic sample. The pH sensitive surfactant may change the surface tension of the droplet responsive to amplification of the nucleic acid molecule. The droplet may be sorted into one of a plurality of sorting lanes based on the surface tension of the droplet, where a sorting lane among the plurality of sorting lanes is associated with droplets including the amplified nucleic acid molecule. A determination of whether the droplet includes the amplified nucleic acid molecule may be performed by detecting passage of the droplet in one of the plurality of sorting lanes.

A system and method for processing and detecting nucleic acids from a set of biological samples, comprising: a capture plate and a capture plate module configured to facilitate binding of nucleic acids within the set of biological samples to magnetic beads; a molecular diagnostic module configured to receive nucleic acids bound to magnetic beads, isolate nucleic acids, and analyze nucleic acids, comprising a cartridge receiving module, a heating/cooling subsystem and a magnet configured to facilitate isolation of nucleic acids, a valve actuation subsystem configured to control fluid flow through a microfluidic cartridge for processing nucleic acids, and an optical subsystem for analysis of nucleic acids; a fluid handling system configured to deliver samples and reagents to components of the system to facilitate molecular diagnostic protocols; and an assay strip configured to combine nucleic acid samples with molecular diagnostic reagents for analysis of nucleic acids.

A reaction processor is provided with a reaction processing vessel having a channel, a liquid feeding system, a temperature control system, and a fluorescence detector, and a CPU for controlling the liquid feeding system. When a sample moves from a low temperature region to a high temperature region, the CPU instructs the liquid feeding system to stop the sample when a predetermined first waiting time has passed from the time when the passage of the sample through a fluorescence detection region is detected by the fluorescence detector. When the sample moves from the high temperature region to the low temperature region, the CPU instructs the liquid feeding system to stop the sample when a predetermined second waiting time, which is set independently of the first waiting time, has passed from the time when the passage of the sample through the fluorescence detection region is detected by the fluorescence detector.

Air-matrix digital microfluidics (DMF) apparatuses and methods of using them to prevent or limit evaporation and surface fouling of the DMF apparatus. In particular, described herein are air-matrix DMF apparatuses and methods of using them including thermally controllable regions with a wax material that may be used to selectively encapsulate a reaction droplet in the air gap of the apparatus; additional aqueous droplets may be combined with the encapsulated droplet even after separating from the wax, despite residual wax coating, by merging with an aqueous droplet having a coating of a secondary material (e.g., an oil or other hydrophobic material) that may remove the wax from the droplet and/or allow combining of the droplets.

A reaction processing vessel includes a substrate and a groove-like channel formed on the upper surface of the substrate. The channel includes a high temperature serpiginous channel, a medium temperature serpiginous channel, and a high temperature braking channel and a medium temperature braking channel that are adjacent to the high temperature serpiginous channel and the medium temperature serpiginous channel, respectively. The respective cross-sectional areas of the high temperature braking channel and the medium temperature braking channel are larger than the respective cross-sectional areas of the high temperature serpiginous channel and the medium temperature serpiginous channel, respectively.

A rapid PCR detection device includes a body, a disposable microfluidic unit, a magnetron micro-fluid unit, a linear actuator, a PCR thermal cycling unit and an image recognition unit. The microfluidic unit is made of a transparent material, wherein a transparent film is arranged in the middle of the microfluidic channel and has at least one hole for a micro-fluid to flow in the microfluidic channel. The magnetron micro-fluid unit drives the micro-fluid, so that the micro-fluid is divided into a plurality of droplets each guided to a lower layer of the microfluidic channel. The linear actuator drives the disposable microfluidic unit to an amplification zone. The PCR thermal cycling unit performs PCR thermal cycling in the amplification zone. The image recognition unit illuminates the droplets with a fluorescent light and determines the number of DNA fragments in the droplets according to the detected fluorescent intensity.