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 method for isolating a nanoparticulate in a sample, the method comprising:
a. applying the sample to a device, the device comprising an array of electrodes capable of establishing an AC electrokinetic field region;
b. concentrating cells or other particulate material in the fluid in a first AC electrokinetic field region, wherein the conductivity of the fluid is 100 mS/m or greater;
c. isolating the nanoparticulate in a second AC electrokinetic field region, wherein the second AC electrokinetic field is a second dielectrophoretic high field region; and
d. flushing the concentrated cells and/or other particulate material from the first AC electrokinetic field region.
2. The method of claim 1 , wherein the nanoparticulate comprises high molecular weight nucleic acids, high molecular weight DNA, smaller DNA, aggregated proteins, cellular debris, RNA, viruses or combinations thereof.
3. The method of claim 1 , wherein detection of the nanoparticulate provides information useful for cancer diagnosis, cancer prognosis or treatment response in a patient.
4. The method of claim 3 , wherein detection of the nanoparticulate is part of a medical therapeutic or diagnostic procedure.
5. The method of claim 1 , further comprising eluting the nanoparticulate isolated in the second dielectrophoretic high field region for further characterization.
6. The method of claim 1 , wherein the sample is whole blood, serum, plasma, cerebrospinal fluid, body tissue, urine or saliva.
7. The method of claim 1 , wherein the sample is blood.
8. The method of claim 1 , wherein the AC electrokinetic field is produced using an alternating current having a voltage of 1 volt to 40 volts peak-peak, and/or a frequency of 5 Hz to 5,000,000 Hz and duty cycles from 5% to 50%.
9. The method of claim 1 , wherein the conductivity of the fluid is 500 mS/m or greater.
10. The method of claim 1 , wherein the array of electrodes is spin-coated with a hydrogel having a thickness between about 0.1 microns and 1 micron.
11. The method of claim 10 , wherein the hydrogel comprises two or more layers of a synthetic polymer.
12. The method of claim 10 , wherein the hydrogel has a viscosity between about 0.5 cP to about 5 cP prior to spin-coating.
13. The method of claim 10 , wherein the hydrogel has a conductivity between about 0.1 S/m to about 1.0 S/m.
14. The method of claim 1 , wherein the array of electrodes comprises a wavy line configuration, wherein the configuration comprises a repeating unit comprising the shape of a pair of dots connected by linker, wherein the linker tapers inward toward 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.
15. The method of claim 1 , wherein the array of electrodes comprises a passivation layer with a relative electrical permittivity from about 2.0 to about 4.0.
16. A method for isolating a nanoparticulate in a sample, the method comprising:
a. applying the sample to a device, the device comprising an array of electrodes capable of establishing an AC electrokinetic field region;
b. concentrating cells or other particulate material in the fluid in a first AC electrokinetic field region, wherein the conductivity of the fluid is 100 mS/m or greater;
c. isolating the nanoparticulate in a second AC electrokinetic field region, wherein the second AC electrokinetic field is a second dielectrophoretic high field region;
d. flushing the concentrated cells and/or other particulate material from the first AC electrokinetic field region; and
e. eluting the bound nanoparticulate in the second dielectrophoretic high field region.
17. The method of claim 16 , wherein the nanoparticulate comprises high molecular weight nucleic acids, high molecular weight DNA, smaller DNA, aggregated proteins, cellular debris, RNA, viruses or combinations thereof.
18. The method of claim 16 , wherein detection of the nanoparticulate provides information useful for cancer diagnosis, cancer prognosis or treatment response in a patient.
19. The method of claim 18 , wherein detection of the nanoparticulate is part of a medical therapeutic or diagnostic procedure.
20. The method of claim 16 , wherein the sample is whole blood, serum, plasma, cerebrospinal fluid, body tissue, urine or saliva.
21. The method of claim 20 , wherein the sample is blood.
22. The method of claim 16 , wherein the AC electrokinetic field is produced using an alternating current having a voltage of 1 volt to 40 volts peak-peak, and/or a frequency of 5 Hz to 5,000,000 Hz and duty cycles from 5% to 50%.