Nucleic acid sample preparation
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.
1. A device for isolating DNA from a fluid comprising cells, the device comprising:
a. a first array of electrodes capable of establishing a first AC electrokinetic field region, wherein the first AC electrokinetic field region is a first dielectrophoretic low field region capable of concentrating and lysing a plurality of cells in a conductivity of fluid of 100 mS/m or more;
b. a second array of electrodes capable of establishing a second AC electrokinetic field, wherein the second AC eletrokinetic field is a dieletrophoretic high field region capable of concentrating and isolating nucleic acid in a conductivity of fluid of 100 mS/m or more;
c. a reservoir comprising a fluid and a device for flushing residual and lysed cellular material from the first array of electrodes; and
d. a module capable of performing sequencing.
2. The device of claim 1 , wherein the first and second array of electrodes are configured to be selectively energized to establish a dielectrophoretic high field and dielectrophoretic low field regions.
3. The device of claim 1 , wherein the first array of electrodes and the second array of electrodes is in a wavy or nonlinear line configuration, wherein the configuration comprises a repeating unit comprising the shape of a pair of dots connected by a linker, wherein the dots and linker define the boundaries of the electrode, wherein the linker tapers inward towards or at the midpoint between the pair of dots, wherein the diameters of the dots are the widest points along the length of the repeating unit, wherein the edge to edge distance between a parallel set of repeating units is equidistant, or roughly equidistant.
4. The device of claim 1 , wherein nucleic acid is isolated and sequencing is performed in a single chamber.
5. The device of claim 1 , wherein nucleic acid is isolated and sequencing is performed in multiple regions of a single chamber.
6. The device of claim 1 , further comprising using at least one of an elution tube, a chamber and a reservoir to perform sequencing.
7. The device of claim 1 , wherein sequencing of the nucleic acid is performed in a serpentine microchannel comprising a plurality of temperature zones.
8. The device of claim 1 , wherein sequencing is performed in aqueous droplets entrapped in immiscible fluids.
9. The device of claim 1 , wherein the device comprises a module capable of thermocycling and amplifying nucleic acids.
10. The device of claim 1 , wherein the device comprises a surface contacting or proximal to the electrodes, wherein the surface is functionalized with biological ligands that are capable of selectively capturing biomolecules.
11. The device of any of claim 1 , wherein the array of electrodes is coated with a hydrogel.
12. The device of claim 11 , wherein the hydrogel comprises two or more layers of a synthetic polymer.
13. The device of claim 11 , wherein the hydrogel is spin-coated onto the electrodes.
14. The device of claim 11 , wherein the hydrogel has a viscosity between about 0.5 cP to about 5 cP prior to spin-coating.
15. The device of any claim 11 , wherein the hydrogel has a thickness between about 0.01 microns and 1 micron.
16. The device of any of claim 11 , wherein the hydrogel has a conductivity between about 0.1 S/m to about 1.0 S/m.
17. The device of claim 1 , wherein the module capable of performing sequencing is a module capable of performing Sanger sequencing.
18. The device of claim 17 , wherein the module capable of performing Sanger sequencing comprises a module capable of capillary electrophoresis, a module capable of multi-color fluorescence detection, or a combination thereof.
19. The device of claim 1 , wherein the module capable of performing sequencing is a module capable of performing next generation sequencing.
20. The device of claim 19 , wherein the module capable of performing next-generation sequencing is capable of performing pyrosequencing, ion semiconductor sequencing, polony sequencing, sequencing by ligation, DNA nanoball sequencing, or single molecule sequencing.