Nucleic acid sample preparation
Inventors
Krishnan, Rajaram • CHARLOT, David • Tu, Eugene • MCCANNA, James • Kumosa, Lucas • Swanson, Paul • Turner, Robert • Yang, Kai • DOBROVOLSKAYA, Irina • Liu, David
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9827565-B2
Publication Date
2017-11-28
Expiration Date
Abstract
The present invention includes methods, devices and systems for isolating a nucleic acid from a fluid comprising cells. In various aspects, the methods, devices and systems may allow for a rapid procedure that requires a minimal amount of material and/or results in high purity nucleic acid isolated from complex fluids such as blood or environmental samples.
Core Innovation
The invention provides a device for isolating DNA from a fluid comprising cells. The device establishes a first AC electrokinetic field region that is a dielectrophoretic low field region capable of concentrating and lysing a plurality of cells in a fluid having a conductivity of 100 mS/m or more, and a second AC electrokinetic field that forms a dielectrophoretic high field region capable of concentrating and isolating nucleic acid in a fluid having a conductivity of 100 mS/m or more.
The device includes a reservoir comprising a fluid and a device for flushing residual and lysed cellular material from the first array of electrodes. This flushing feature is positioned to remove residual and lysed cellular material associated with the concentration and lysis occurring in the first dielectrophoretic low field region.
In the described embodiments, the electrode arrays can be selectively energized to form dielectrophoretic high-field and dielectrophoretic low-field regions. The system is integrated with sequencing capability, including sequencing modules that can perform Sanger sequencing, and the document also describes functionalized capture surfaces and hydrogel coatings near the electrodes.
Claims Coverage
The independent claim provides a DNA isolation device with two dielectrophoretic AC electrokinetic field regions, a low field for concentrating and lysing cells and a high field for concentrating and isolating nucleic acid, in fluids having conductivity of 100 mS/m or more, together with flushing of residual and lysed material and a sequencing module. Dependent claims refine electrode energization and add functionalized capture surfaces, hydrogel coatings, and sequencing modality features.
Two-stage dielectrophoretic low-field and high-field AC electrokinetic regions for conductivity-defined cell lysis and nucleic-acid isolation
A first array of electrodes establishes a dielectrophoretic low field region to concentrate and lyse cells, and a second array of electrodes establishes a dielectrophoretic high field region to concentrate and isolate nucleic acid, with both regions operated in fluids having conductivity of 100 mS/m or more.
Flushing residual and lysed cellular material from the first array of electrodes
The device includes a reservoir comprising a fluid and a device for flushing residual and lysed cellular material from the first array of electrodes.
Sequencing-capable module integrated with the DNA isolation device
The device includes a module capable of performing sequencing.
Selective energization of first and second electrode arrays to establish dielectrophoretic regions
The first and second electrode arrays are configured to be selectively energized to establish dielectrophoretic high-field and dielectrophoretic low-field regions.
Functionalized capture surface near or contacting the electrodes for selective biomolecule capture
A surface near or contacting the electrodes is functionalized with biological ligands configured to selectively capture biomolecules.
Hydrogel coating near the electrodes
The device includes a hydrogel coating near the electrodes.
Sanger-sequencing sequencing module
The device includes a sequencing-capable module specifically configured to perform Sanger sequencing.
Serpentine microchannel with multiple temperature zones for sequencing
Nucleic acid sequencing is performed in a serpentine microchannel having multiple temperature zones.
The claims center on AC electrokinetic electrode arrays that create a dielectrophoretic low-field region for concentrating and lysing cells and a dielectrophoretic high-field region for concentrating and isolating nucleic acids at conductivity of 100 mS/m or more, with flushing of residual and lysed material. The claims also specify integration with sequencing capability and further features including selectable energization, functionalized capture surfaces, hydrogel coatings, Sanger sequencing, and serpentine microchannel temperature zones.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
Interested in licensing this patent?