摘要:
The invention provides pyrosequencing-based methods of analyzing and synthesizing DNA, including methods of DNA error correction, determining DNA size distribution, screening for nucleotide repeat disorders such as fragile X syndrome, determining size distribution and bias in a DNA library, and determining pyrosequencing read length. The methods include on-bench protocols as well as droplet-based protocols that may be conducted on a droplet actuator.
摘要:
A method, comprising: generating an output voltage by a power supply; storing charge produced by the output voltage; superimposing an excitation signal onto the output voltage to produce a superimposed signal; connecting the superimposed signal to an electrode in a droplet actuator; suppressing the output voltage from the power supply when detecting an impedance at the electrode; supplying the charge to the droplet actuator to activate the electrode during the impedance; and measuring the impedance produced by the excitation signal while the output voltage is suppressed, wherein the impedance indicates presence of liquid at the electrode. Superimposing the excitation signal may comprise adding the excitation signal to the output voltage. Suppressing the output voltage may comprise at least one of stopping the output voltage generated by the power supply, disabling a switching action of the power supply, and disabling the power supply during the impedance measurement.
摘要:
The invention provides droplet actuator assemblies and systems and methods of manufacturing the droplet actuator assemblies. In certain embodiments, two-piece enclosures are used to form a droplet actuator assembly that houses a droplet operations substrate. In certain other embodiments, one-piece enclosures are used to form a droplet actuator assembly that houses a droplet operations substrate. In the plastic injection molding process for forming substrates of the droplet actuator assemblies of the present invention may utilize insert molding (or overmolding) processes for forming a gasket in at least one substrate, thereby avoiding the need for providing and installing a separate gasket component. Further, the droplet actuator assemblies may include features that allow ultrasonic welding processes to be used for bonding substrates together. The manufacturing systems of the present invention for fabricating the droplet actuator assemblies may utilize continuous flow reel-to-reel manufacturing processes.
摘要:
Devices, configurations and methods for improving absorbance detection are provided. For example, methods and devices are provided for determining the absorbance of a droplet, e.g., a droplet on a droplet actuator, by providing an elongated light path through the droplet.
摘要:
The invention relates to certain novel approaches to reducing or eliminating the movement of contaminants from one droplet to another on a droplet actuator via liquid filler fluid. In one application, droplet actuators are used to conduct genetic analysis using polymerase chain reaction (PCR) techniques. The invention addresses the need for improved methods of performing PCR on a droplet actuator that provide for optimum amplification and detection of a sample target.
摘要:
The invention relates to certain novel approaches to reducing or eliminating the movement of contaminants from one droplet to another on a droplet actuator via liquid filler fluid. In one application, droplet actuators are used to conduct genetic analysis using polymerase chain reaction (PCR) techniques. The invention addresses the need for improved methods of performing PCR on a droplet actuator that provide for optimum amplification and detection of a sample target.
摘要:
A method of assembling a droplet actuator assembly (1400) comprises: attaching to a mount structure (1410) a substrate (1434) including an electrode emitting an electric field (1438) affecting a droplet present in the droplet actuator assembly and a magnet assembly (1420) configured to accommodate a magnet (1428) emitting a magnetic field affecting a magnetically responsive bead present in the droplet actuator assembly. At least one of the substrate (1434) and the magnet assembly (1420) are removably attached to the mount structure (1410). Attaching the substrate (1434) may further comprise selecting the substrate from among a plurality of substrates (1434) configured to attach to the mount structure (1410) and including different electric field emission characteristics, and/or selecting the magnet assembly (1420) from among a plurality of magnetic assemblies (1420), each magnet assembly configured to attach to the mount structure (1410) and including different magnet properties.
摘要:
A method of preparing a nucleic acid library in droplets in contact with oil, including: (a) blunt-ending nucleic acid fragments in a droplet in the oil to yield blunt-ended nucleic acid fragments; (b) phosphorylating the blunt-ended nucleic acid fragments in a droplet in the oil to yield phosphorylated nucleic acid fragments; coupling A-tails to the phosphorylated nucleic acid fragments in a droplet in the oil to yield A-tailed nucleic acid fragments; and (d) coupling nucleic acid adapters to the A-tailed nucleic acid fragments in a droplet in the oil to yield the nucleic acid library comprising adapter-ligated nucleic acid fragments.