Apparatus and method for purification of nucleic acids by phase separation using laser and beads

• US 20060094051A1
• Filed: 11/02/2005
• Published: 05/04/2006
• Est. Priority Date: 11/03/2004
• Status: Active Grant

• Current Assignee: Samsung Electronics Co. Ltd.
• Original Assignee: Samsung Electronics Co. Ltd.
• Inventors: Lee, Jeong-gun, Kwon, Young-nam, Kim, Young-a
• Granted Patent: US 7,429,470 B2
• Time in Patent Office: Days
• US Class Current: 435/6.11
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0633 with moving parts
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: G01N 35/0098 involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

An apparatus and method for purification of nucleic acids of cells or viruses are provided. The nucleic acid purification apparatus includes: a cell lysis capillary having a sample inlet through which samples and magnetic beads are introduced; a vibrator attached to the capillary and mixing the samples and the magnetic beads in the capillary; a laser generator attached to the capillary and supplying a laser to the capillary; and a magnetic force generator attached to the capillary and fixing the magnetic beads to a capillary wall. According to the method and apparatus, PCR yield can be increased since PCR inhibitors can be readily removed by means of a phase separation in a capillary. The use of an electromagnet ensures the removal of the PCR inhibitors. In addition, since cell lysis and DNA purification process can be simultaneously performed, LOC steps can be reduced.

Claims

1. A nucleic acid purification apparatus of cells or viruses comprising:

a cell lysis capillary having a sample inlet through which samples and magnetic beads are introduced;

a vibrator attached to the capillary and mixing the samples and the magnetic beads in the capillary;

a laser generator attached to the capillary and supplying a laser to the capillary; and

a magnetic force generator attached to the capillary and fixing the magnetic beads to a capillary wall.

2. The apparatus of claim 1, wherein the vibrator is selected from the group consisting of sonicators, vibrators using a magnetic field, vibrators using an electric field, mechanical vibrators and piezoelectric materials.

3. The apparatus of claim 1, wherein the magnetic force generator is located above a laser pathway and is an electromagnet which is turned on when the magnetic beads in the cell lysis capillary are boiled.

4. The apparatus of claim 1, further comprising a DNA amplification chamber connected to the cell lysis capillary through a channel which is opened or closed by a valve.

5. A method of purifying nucleic acids using the nucleic acid purification apparatus of claim 1, the method comprising:

injecting a solution containing cells or viruses to a capillary-shaped container containing magnetic beads;

vibrating the magnetic beads;

irradiating a laser beam onto the magnetic beads to disrupt the cells or viruses and binding compounds in the resulting cell or virus lysate to the magnetic beads;

fixing the magnetic beads, to which the compounds in the cell or virus lysate are bound, to a capillary-shaped container wall by means of a magnetic force generator; and

obtaining a solution without the magnetic bead.

6. A method of continuously performing purification and amplification of nucleic acids using the nucleic acid purification apparatus of claim 4, the method comprising:

injecting a solution containing cells or viruses to a capillary-shaped container containing magnetic beads;

vibrating the magnetic beads;

irradiating a laser beam onto the magnetic beads to disrupt the cells or viruses and binding compounds in the resulting cell or virus lysate to the magnetic beads;

fixing the magnetic beads, to which the compounds in the cell or virus lysate are bound, to a capillary-shaped container wall by means of a magnetic force generator; and

obtaining a solution without the magnetic bead and transferring the resulting solution to a amplification chamber through a channel connecting the container and the amplification chamber to perform amplification.

7. The method of claim 5, wherein the laser comprises a pulse laser or continuous wave (CW) laser.

8. The method of claim 7, wherein the pulse laser is 1 mJ/pulse or more and the CW laser has a power of 10 mW or more.

9. The method of claim 8, wherein the pulse laser is 32 mJ/pulse or more and the CW laser has a power of 10 W or more.

10. The method of claim 5, wherein the laser beam is generated in a wavelength range of 400 m or more.

11. The method of claim 10, wherein the laser beam is generated in a wavelength range of from 750 nm to 1300 nm.

12. The method of claim 10, wherein the laser is generated in one or more wavelength ranges.

13. The method of claim 5, wherein the size of the magnetic beads is from 50 nm to 1,000 μ

m.

14. The method of claim 13, wherein the size of the magnetic beads is 1-50 μ

m.

15. The method of claim 13, wherein the magnetic beads is a mixture of beads having two or more sizes.

16. The method of claim 5, wherein the container has a ratio of diameter to length ranging from 1:

2 to 1;

50.

17. The method of claim 16, wherein the container has a diameter ranging from 1 nm to 5 mm.

18. The method of claim 5, wherein the container is selected from the group consisting of polymers, organic materials, silicon, glass and metals.

19. The method of claim 5, wherein the magnetic beads comprise at least one material selected from the group consisting of ferromagnetic Fe, Ni, Cr, and oxides thereof.

20. The method of claim 5, wherein the magnetic beads are polymers, organic materials, silicon or glass coated with a ferromagnetic metal.

21. The method of claim 5, wherein the magnetic beads have a negatively charged surface.

22. The method of claim 5 or wherein the solution is selected from the group consisting of saliva, urine, blood, serum and cell cultures.

23. The method of claim 6, wherein the laser comprises a pulse laser or continuous wave (CW) laser.

24. The method of claim 6, wherein the laser beam is generated in a wavelength range of 400 nm or more.

25. The method of claim 6, wherein the size of the magnetic beads is from 50 nm to 1,000 μ

m.

26. The method of claim 6, wherein the container has a ratio of diameter to length ranging from 1:

2 to 1;

50.

27. The method of claim 6, wherein the container is selected from the group consisting of polymers, organic materials, silicon, glass and metals.

28. The method of claim 6, wherein the magnetic beads comprise at least one material selected from the group consisting of ferromagnetic Fe, Ni, Cr, and oxides thereof.

29. The method of claim 6, wherein the magnetic beads are polymers, organic materials, silicon or glass coated with a ferromagnetic metal.

30. The method of claim 6, wherein the magnetic beads have a negatively charged surface.

31. The method of claim 6, wherein the solution is selected from the group consisting of saliva, urine, blood, serum and cell cultures.