Method and apparatus for isolating and purifying nucleic acid using a single surface

• US 7,718,371 B2
• Filed: 03/29/2007
• Issued: 05/18/2010
• Est. Priority Date: 09/25/2006
• Status: Active Grant

• Current Assignee: Samsung Electronics Co. Ltd.
• Original Assignee: Samsung Electronics Co. Ltd.
• Inventors: Yoo, Chang eun, Jeong, Sung-young, Kim, Young-rok
• Primary Examiner(s): Riley; Jezia
• Application Number:
  US11/693,259
• Publication Number:
  US 20080118972A1
• Time in Patent Office: 1,146 Days
• Field of Search: 435/5, 435/6, 435/7.1, 435/7.2, 536/23.1, 422/61, 422/68.1, 424/489, 424/490
• US Class Current: 435/6.1
• CPC Class Codes: B01D 15/1807 using counter-currents, e.g. fluidised beds
• CPC Class Codes: B01J 20/28009 Magnetic properties
• CPC Class Codes: B01J 20/3204 Inorganic carriers, supports or substrates
• CPC Class Codes: B01J 20/321 consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3212 consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3219 involving a particular spacer or linking group, e.g. for attaching an active group
• CPC Class Codes: B01J 20/327 Polymers obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: C08F 20/02 Monocarboxylic acids having less than ten carbon atoms, Derivatives thereof
• CPC Class Codes: C08F 26/06 by a heterocyclic ring containing nitrogen
• CPC Class Codes: C12N 1/066 by physical methods
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves

Abstract

Provided is a method of isolating nucleic acid from cells using a single surface, wherein a compound represented by Formula 1 is bound to the surface. Also provided are an apparatus for isolation of nucleic acids, and a bead for isolating nucleic acids.

Claims

1. A method of isolating nucleic acid from cells, the method comprising:

mixing a sample containing cells with a solution comprising beads dispersed in a binding buffer to bind the cells to the beads;

separating the beads having the cells bound thereto from the binding buffer and then washing the beads with a wash buffer;

disrupting the bound cells to release nucleic acid to bind to the beads; and

eluting bound nucleic acid from the beads using an elution buffer,

wherein a compound represented by Formula 1 below is bound to the surface of the beads;

2. The method of claim 1, wherein the disrupting of the cells is performed by heating or by laser-irradiation.

3. The method of claim 2, wherein the laser is a pulse laser or a continuous wave (CW) laser.

4. The method of claim 3, wherein the pulse laser has a power of at least about 3 mJ/pulse, and the CW laser has a power of at least about 100 mW.

5. The method of claim 3, wherein the wavelength of the laser beam is at least about 400 nm.

6. The method of claim 1, wherein the binding buffer has a pH of about 3 to about 5.

7. The method of claim 1, wherein the elution buffer has a pH of about 8 to about 10.

8. The method of claim 1, wherein eluting bound nucleic acid from the beads is performed simultaneously with irradiating the beads with a laser or heating.

9. The method of claim 1, wherein the beads are magnetic beads, silica beads, polystyrene beads, glass beads, or cellulose beads.

10. The method of claim 1, wherein R₁₀ is a pyridinyl group or an imidazolyl group.

11. The method of claim 1, wherein R₁ and R₅ are each a hydroxyl group.

12. The method of claim 1, wherein the sample is saliva, urine, blood, serum, or a cell culture.