Search

US-20260125668-A1 - DEVICES, SYSTEMS, AND METHODS RELATING TO MICROFLUIDIC PURIFICATION OF NUCLEIC ACIDS

US20260125668A1US 20260125668 A1US20260125668 A1US 20260125668A1US-20260125668-A1

Abstract

Described herein are devices, systems, and methods related to isolation and/or detection and/or trapping of nucleic acids utilizing microfluidics. In an embodiment, described herein is a microfluidic trap. In embodiments, a microfluidic trap can comprise an inlet region having an injection port, a first extraction port, and a first electrode; an outlet region having an outlet port and a second electrode; and a microfluidic channel providing fluidic communication between the inlet region and outlet region. In certain aspects, nucleic acids that are trapped comprise genomic DNA.

Inventors

  • Jason Edward Butler
  • Anthony J. Ladd
  • Jiayi Wang

Assignees

  • UNIVERSITY OF FLORIDA RESEARCH FOUNDATION, INC.

Dates

Publication Date
20260507
Application Date
20251031

Claims (20)

  1. 1 . A microfluidic trap, comprising: an inlet region having an injection port, a first extraction port, and a first electrode; an outlet region having an outlet port and a second electrode; and a microfluidic channel providing fluidic communication between the inlet region and outlet region.
  2. 2 . The microfluidic trap of claim 1 , wherein the first electrode and the first extraction port are in close proximity and separated from the injection port by an inlet distance.
  3. 3 . The microfluidic trap of claim 1 , wherein the second electrode and the outlet port are separated by an outlet distance.
  4. 4 . The microfluidic trap of claim 1 , wherein the injection port, extraction port, and outlet port are in line with a longitudinal axis of the microfluidic channel.
  5. 5 . The microfluidic trap of claim 1 , wherein the inlet region further comprises one or more reservoirs adjacent to the longitudinal axis of the microfluidic channel, wherein the first electrode is within one of the one or more reservoirs and the second electrode is in line with the longitudinal axis of the microfluidic channel.
  6. 6 . The microfluidic trap of claim 1 , wherein the first electrode and the second electrode comprise a noble metal, one or more metal alloys, or a combination of any thereof.
  7. 7 . The microfluidic trap of claim 1 , wherein the first electrode and second electrode are coated with a charge neutral polymer having a sufficient pore size to prevent passage of genomic DNA or other nucleic acids through the pores.
  8. 8 . The microfluidic trap of claim 1 , wherein the inlet region, outlet region, microfluidic channel, or any combination of any thereof are laser etched in acrylic.
  9. 9 . The microfluidic trap of claim 1 , wherein the inlet region, outlet region, microfluidic channel, or any combination of any thereof comprise glass, silicon, polydimethylsiloxane (PDMS), paper, a thermoplastic material, or a combination of any thereof.
  10. 10 . A system, comprising: a microfluidic trap of claim 1 ; a voltage generator; one or more fluid flow valves; and a fluid flow generator.
  11. 11 . The system of claim 10 , further comprising: a fluid source in fluidic communication with the inlet region of the microfluidic trap.
  12. 12 . The system of claim 10 , further comprising: a fluid collection device in fluidic communication with the outlet region of the microfluidic trap.
  13. 13 . The system of claim 10 , further comprising: a flow valve in fluidic communication with the fluid source.
  14. 14 . The system of claim 10 , wherein the fluid flow generator is an active or passive fluid flow generator.
  15. 15 . The system of claim 14 , wherein the active fluid flow generator is a fluidic pump.
  16. 16 . The system of claim 14 , wherein the passive fluid flow generator is gravitational flow created by a difference in height between the fluid source and the fluid collection device.
  17. 17 . A method, comprising: establishing a fluid flow in the system of claim 10 ; providing a first electric field across the longitudinal axis of the microfluid channel with the voltage generator; injecting a biological sample comprising genomic DNA into the injection port of the system; waiting a first period of time; stopping the fluid flow; removing the first electric field; providing a second electric field across the first and second electrodes of the microfluidic trap with the voltage generator; waiting a second period of time; collecting the remaining nucleic acids from the extraction port of the microfluidic trap.
  18. 18 . The method of claim 17 , wherein the biological sample comprises cell lysate comprising genomic DNA.
  19. 19 . The method of claim 17 , wherein the polarity of the first electric field is such that the polarity attracts a polyelectrolyte or nucleotide to the inlet region.
  20. 20 . The method of claim 17 , wherein the polarity of the second electric field is such that the polarity attracts a polyelectrolyte or nucleotide to the inlet region.

Description

CROSS-REFERENCE TO RELATED APPLICATION[S] This application claims priority to, and the benefit of, U.S. Provisional Patent Application Ser. No. 63/715,486, entitled “MICROFLUIDIC DNA PURIFICATION PROCESS” and filed on Nov. 1, 2024, the entire contents of which are incorporated herein by reference as if set forth in its entirety. This application claims priority to, and the benefit of, U.S. Provisional Patent Application Ser. No. 63/743,442, entitled “MICROFLUIDIC DNA PURIFICATION PROCESS” and filed on Jan. 9, 2025, the entire contents of which are incorporated herein by reference as if set forth in its entirety. This application claims priority to, and the benefit of, U.S. Provisional Patent Application Ser. No. 63/807,946, entitled “DEVICES, SYSTEMS, AND METHODS RELATING TO MICROFLUIDIC PURIFICATION OF NUCLEIC ACIDS” and filed on May 19, 2025, the entire contents of which are incorporated herein by reference as if set forth in its entirety. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with government support under Grant No(s) 1804302 & 2222688, awarded by the National Science Foundation. The government has certain rights in the invention. SEQUENCE LISTING The instant application contains a Sequence Listing which has been submitted electronically in .XML format and is hereby incorporated by reference in its entirety. Said. XML copy, created on Oct. 29, 2025, is named “UFPT19570US004_222112_1970_sequence listing.xml” and is 2,932 bytes in size. BACKGROUND Microfluidic processing and analysis of DNA continues to be a vibrant area of research and development due to the reduced sample sizes and possibilities for integrating sample preparation and analysis into a single fully automated device. A cell lysate contains a number of contaminants that must be removed before any analysis of the DNA is possible. These include components native to the biological sample, such as polysaccharides in plant-based samples, or reagents used in the lysing process, such as salts and detergents. In practice, purification is typically performed on relatively large samples (milliliter volumes) using liquid or solid-phase extraction methods, including magnetic beads that bind reversibly to the DNA. However, these methods are time-consuming and labor-intensive, involving multiple pipette transfers and centrifugation steps. The stresses created by centrifuging fragment individual DNA strands, which is undesirable when preparing samples for long-read sequencing. There is a need to address the aforementioned deficiencies and inadequacies accordingly. SUMMARY Described herein are devices, kits, systems, compositions, and methods related to isolation and/or detection and/or trapping of nucleic acids utilizing fluid flow and electric field application within microfluidics. Described herein are microfluidic traps, systems, and methods of use. In embodiments, described herein is a microfluidic trap, comprising an inlet region having an injection port, a first extraction port, and a first electrode; an outlet region having an outlet port and a second electrode; and a microfluidic channel providing fluidic communication between the inlet region and outlet region. In certain aspects, there may be electrodes in the microfluidic device and outside of the microfluidic device. The polyelectrolyte trapping part of the process can be driven by the external electrodes, but can also be driven by the internal electrodes. The internal electrodes must be used for DNA collection in order to avoid reintroducing contaminants. In embodiments, the first electrode and the first extraction port are in close proximity and separated from the injection port by an inlet distance. In embodiments, the second electrode and the outlet port are separated by an outlet distance. In embodiments, the injection port, extraction port, and outlet port are in line with the longitudinal axis of the microfluidic channel. In embodiments, the inlet region further comprises one or more reservoirs adjacent to the longitudinal axis of the microfluidic channel, wherein the first electrode is within one of the one or more reservoirs and the second electrode is in line with the longitudinal axis of the microfluidic channel. In certain aspects, the first electrode can be positioned in a secondary, side channel or area which is connected to the inlet, or in line with the inlet. In certain aspects, an inlet electrode would need to be near to the extraction port. In certain aspects, the second electrode probably is inline, or at least not offset very far, from the inlet or microfluidic channel. In embodiments, the first electrode and the second electrode comprise a noble metal, one or more metal alloys, or a combination of any thereof. In embodiments, the first electrode and second electrode can be coated with a charge neutral polymer having a sufficient pore size to prevent passage of genomic DNA or other nucleic acids through the pores. In embodimen