Apparatus for and method of purifying nucleic acids by different laser absorption of beads

• US 20060110725A1
• Filed: 07/26/2005
• Published: 05/25/2006
• Est. Priority Date: 11/25/2004
• Status: Abandoned Application

• Current Assignee: Samsung Electronics Co. Ltd.
• Original Assignee: Samsung Electronics Co. Ltd.
• Inventors: Lee, Jeong-gun, Kwon, Young-nam, Lee, Myo-yong, Cho, Yeon-ja, Yoo, Shin-i, Kim, Young-a
• Application Number:
  US11/190,169
• Publication Number:
  US 20060110725A1
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• 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 2300/0681 Filter
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• 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

Abstract

An apparatus for and method of purifying 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, magnetic beads, and a solid support are introduced; a vibrator attached to the capillary and mixing the samples, magnetic beads, and solid support in the capillary; a laser generator attached to the capillary and irradiating a laser beam onto the capillary; a magnetic force generator attached to the capillary and fixing the magnetic beads to a capillary wall; a waste chamber attached to the capillary and discharging a lysate; an elution buffer chamber attached to the capillary and eluting nucleic acids from the solid support having nucleic acids bound thereto; and a neutralization buffer chamber attached to the capillary and supplying a neutralization buffer for neutralizing the eluted nucleic acid solution. According to the apparatus and method, PCR inhibitors can be removed to increase PCR yield and nucleic acids can be purified using a silicon substrate or silica beads. Thus, the apparatus and method can be applied to LOC fabrication.

Claims

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

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

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

a laser generator attached to the capillary and irradiating a laser beam onto the capillary;

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

a waste chamber attached to the capillary and discharging a lysate;

an elution buffer chamber attached to the capillary and eluting nucleic acids from the solid support having nucleic acids bound thereto; and

a neutralization buffer chamber attached to the capillary and supplying a neutralization buffer for neutralizing an eluted nucleic acid solution.

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, and mechanical vibrators.

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. The apparatus of claim 1, further comprising a membrane which is located in a channel disposed between the cell lysis capillary and the DNA amplification chamber and filters the solid support.

6. The apparatus of claim 1, further comprising a washing buffer chamber attached to the capillary and washing the solid support having nucleic acids bound thereto.

7. 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 in a capillary-shaped container containing magnetic beads and a solid support;

operating a vibrator to mix the solution, the magnetic beads and the solid support;

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 and binding nucleic acids in the lysate to the solid support;

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;

discharging the lysate which contains no magnetic bead; and

eluting nucleic acids from the solid support and neutralizing them.

8. 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 and a solid support;

operating a vibrator to mix the solution, the magnetic beads, and the solid support;

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 and binding nucleic acids in the lysate to the solid support;

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;

discharging the lysate which contains no magnetic bead;

eluting the nucleic acids from the solid support and neutralizing an eluted nucleic acid solution; and

obtaining a solution that contains nucleic acids and transferring the resulting solution to a amplification chamber through a channel connecting the container and the amplification chamber to perform amplification.

9. The method of claim 7, further comprising washing the solid support to which nucleic acids are bound and discharging a washing solution after discharging the lysate.

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

11. The method of claim 10, wherein the pulse laser is 1 mJ/pulse to 1 J/pulse and the CW laser has a power of 10 mW to 300 W.

12. The method of claim 7, wherein the laser beam is generated in a wavelength range of from 750 nm to 5000 nm.

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

14. The method of claim 7, wherein the size of the magnetic bead is from 50 nm to 1,000 μ

m.

15. The method of claim 14, wherein the magnetic beads are a mixture of beads having two or more sizes.

16. The method of claim 7, wherein the capillary-shaped 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 7, wherein the container is composed of a material selected from the group consisting of polymers, organic materials, silicon, glass and metals.

19. The method of claim 7, 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 7, wherein the magnetic beads are polymers, organic materials, silicon or glass coated with a ferromagnetic metal.

21. The method of claim 7, wherein the magnetic beads have a negatively charged surface.

22. The method of claim 7, wherein the solution is selected from the group consisting of saliva, urine, blood, serum and cell cultures.

23. The method of claim 7, wherein the solid support is selected from the group consisting of silica beads, a silicon substrate, germanium, diamond, quartz, and silicone.

24. The method of claim 23, wherein the size of the silica bead is 50 nm to 1,000 μ

m.

25. The method of claim 24, wherein the silica beads are a mixture of beads having two or more sizes.

26. The method of claim 24, wherein the silica bead is coated with a positively-charged material.

27. The method of claim 23, wherein the silicon substrate is in a pillar form or has silica beads fixed thereto.