DEVICES AND PROCESSES FOR NUCLEIC ACID EXTRACTION

• US 20120237939A1
• Filed: 04/20/2012
• Published: 09/20/2012
• Est. Priority Date: 06/26/2006
• Status: Abandoned Application

• Current Assignee: Blood Cell Storage, Inc.
• Original Assignee: Blood Cell Storage, Inc.
• Inventors: Reed, Michael W., Nanassy, Oliver Z., Haydock, Paul V., Sharma, Nigel Rudra, Bardell, Ronald L., Hargrave, Perry
• Application Number:
  US13/452,689
• Publication Number:
  US 20120237939A1
• Time in Patent Office: Days
• US Class Current: 435/6.12
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0457 passive flow or gravitation
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0622 distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 2400/084 Passive control of flow resistance
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/6851 : Quantitative amplification
• CPC Class Codes: C12Q 2563/173 : staining/intercalating agent, e.g. ethidium bromide
• CPC Class Codes: C12Q 2565/137 : Chromatographic separation
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device

Abstract

Devices, processes, and kits for the extraction of nucleic acids from biological samples are disclosed. The devices comprise a first port, a second port, and a binding chamber intermediate and in fluid communication with the first port and the second port. The binding chamber comprises an unmodified flat glass surface effective for binding a heterogeneous population of nucleic acids. The first port, second port, and binding chamber define a continuous fluid pathway that is essentially free of nucleic acid-specific binding sites.

Claims

1. A device comprising:

(i) a body member having a plurality of external surfaces and fabricated to contain a continuous fluid pathway therethrough, the pathway comprising;

a first port;

a second port; and

a binding channel intermediate and in fluid communication with the first port and the second port, wherein the binding channel is open to one of the external surfaces of the body member; and

(ii) a glass member affixed to said one of the external surfaces of the body member to provide a first unmodified flat glass surface in fluid communication with the binding channel, wherein the binding channel and glass member define a binding chamber effective for binding a heterogeneous population of nucleic acids and wherein the fluid pathway is essentially free of nucleic acid-specific binding sites.

2. The device of claim 1, wherein the fluid pathway further comprises:

a first channel connecting the first port with the binding chamber; and

a second channel connecting the second port with the binding chamber.

3. The device of claim 1 wherein the binding channel is open to a second of the external surfaces of the body member and wherein the device further comprises a second glass member affixed to the second external surface of the body member to provide a second unmodified flat glass surface in fluid communication with the binding channel.

4. The device of claim 1 wherein the first port or the second port comprises a Luer-lock fitting, an O-ring, a gasket, a tubing stub, or an elastomeric septum.

5. The device of claim 1 wherein the binding chamber comprises a serpentine channel.

6. The device of claim 5 wherein the serpentine channel is planar.

7. The device of claim 1 wherein the binding chamber is rectangular in cross-section.

8. The device of claim 1 further comprising a pump in fluid communication with one of the ports.

9. The device of claim 8 further comprising fluid distribution control means in fluid communication with the pump.

10. The device of claim 9 wherein the fluid distribution control means comprises a programmable computer.

11. The device of claim 1 wherein the fluid pathway further comprises:

a distribution channel in fluid communication with the binding channel; and

a plurality of capillary channels in fluid communication with the distribution channel distal to the binding channel.

12. The device of claim 11 further comprising a plurality of assay wells, wherein each of the assay wells is in fluid communication with one of the capillary channels.

13. A process for extracting nucleic acid from a biological sample comprising:

introducing a nucleic acid-containing sample into the binding chamber of the device of claim 1 via one of the ports;

allowing nucleic acid in the sample to bind to the unmodified flat glass surface;

washing the binding chamber to remove contaminants; and

eluting bound nucleic acid from the unmodified flat glass surface.

14. The process of claim 13 further comprising lysing a cell sample to prepare the nucleic acid-containing sample.

15. The process of claim 13 wherein the nucleic acid is DNA.

16. The process of claim 13 wherein bound nucleic acid is eluted with a buffer containing a fluorescent compound that exhibits a change in fluorescence intensity in the presence of nucleic acids.

17. The process of claim 13 comprising the additional step of amplifying the eluted nucleic acid.

18. The process of claim 13 further comprising drying the washed binding chamber prior to eluting the bound nucleic acid.

19. A process for extracting nucleic acid from a biological sample comprising:

introducing a nucleic acid-containing sample into the binding chamber of the device of claim 1 via one of the ports;

allowing nucleic acid in the sample to bind to the unmodified flat glass surface;

washing the binding chamber to remove contaminants; and

drying the washed binding chamber.

20. The process of claim 19 wherein the nucleic acid is RNA.

21. The process of claim 19 wherein the nucleic acid-containing sample comprises ethanol at a concentration of 10% to 60%.

22. The process of claim 21 wherein the ethanol concentration is 20% to 50%.

23. The process of claim 19 wherein the nucleic acid-containing sample comprises a proteolytic enzyme.

24. The process of claim 19 wherein the nucleic acid is allowed to bind to the unmodified flat glass surface for a period of from five minutes to thirty minutes.

25. A kit comprising:

the device of claim 1; and

a buffer in a sealed container, wherein the buffer is a lysis buffer, a wash buffer, or an elution buffer.