摘要:
The invention provides methods of conducting a nucleic acid reaction, including methods for performing digital PCR using a "droplet-in-oil" technology. In the methods, the starting sampled is segmented at least partially into a set of sample droplets each containing on average about one or fewer copies of a target nucleic acid. The droplets are passed in a continuous flow of immiscible carrier fluid through a channel that passes through a thermal cycler, whereby the target is amplified. In one implementation, the droplets are about 350 nl each and the number of positively amplified droplets is counted at the near-saturation point.
摘要:
A liquid handling system of a PCR system is instructed to obtain a matrix of samples and reagents for a PCR experiment. A fluid pumping system of the PCR system is instructed to maintain a continuous flow of a transport fluid through a plurality of micro-channels that allows the mixture of the samples and the reagents producing a plurality of mixed sample droplets. One or more post-bridge detection values are received from a post-bridge detection system of the PCR system for each mixed sample droplet to determine if the mixed sample droplet is mixed correctly. A thermocycler of the PCR system is instructed to maintain one or more temperatures for cycling the temperature of the plurality of mixed sample droplets. One or more endpoint detection values are received from an endpoint detection system of the PCR system for each mixed sample droplet to analyze the PCR experiment.
摘要:
Provided herein is a biological detection system and method of use wherein the biological detection system comprises at least one mixer or liquid bridge for combining at least two liquid droplets and an error correction system for detecting whether or not proper mixing or combining of the two component droplets have occurred.
摘要:
A bridge (30) comprises a first inlet port (31) at the end of a capillary, a narrower second inlet port (32) which is an end of a capillary, an outlet port (33) which is an end of a capillary, and a chamber (34) for silicone oil. The oil is density-matched with the reactor droplets such that a neutrally buoyant environment is created within the chamber (34). The oil within the chamber is continuously replenished by the oil separating the reactor droplets. This causes the droplets to assume a stable capillary- suspended spherical form upon entering the chamber (34). The spherical shape grows until large enough to span the gap between the ports, forming an axisymmetric liquid bridge. The introduction of a second droplet from the second inlet port (32) causes the formation of an unstable funicular bridge that quickly ruptures from the, finer, second inlet port (32), and the droplets combine at the liquid bridge (30). In another embodiment, a droplet (55) segments into smaller droplets which bridge the gap between the inlet and outlet ports.
摘要:
A thermal cycling device ( 3 ) device a number of fixed thermal zones ( 11, 12, 13 ) and a fixed conduit ( 10 ) passing through the thermal zones. A controller maintains each thermal zone including its section of conduit ( 10 ) at a constant temperature. A series of droplets flows through the conduit ( 10 ) 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 ( 17 ) and a capillary tube ( 10 ) inserted into the channel. The detection system may perform scans along a direction to detect radiation from a plurality of cycles in a pass.
摘要:
The present invention generally relates methods for analyzing agricultural and/or environmental samples using liquid bridges. In certain embodiments, the invention provides a method for analyzing an agricultural sample for a desired trait including obtaining a gene or gene product from an agricultural sample, in which the gene or gene product is in a first fluid; providing a liquid bridge for mixing the gene or gene product with at least one reagent to form a mixed droplet that is wrapped in an immiscible second fluid; and analyzing the mixed droplet to detect a desired trait of the agricultural sample.
摘要:
The present invention generally relates to devices, systems, and methods for acquiring and/or dispensing a sample without introducing a gas into a microfludic system, such as a liquid bridge system. An exemplary embodiment provides a sampling device including an outer sheath; a plurality of tubes within the sheath, in which at least one of the tubes acquires a sample, and at least one tubes expels a fluid that is immiscible with the sample, in which the at least one tube that acquires the sample is extendable beyond a distal end of the sheath and retractable to within the sheath; and a valve connected to a distal portion of the sheath, in which the valve opens when the tube extends beyond the distal end and closes when the tube retracts to within the sheath.