Instrument and method for nucleic acid isolation

• US 20070269829A1
• Filed: 05/17/2007
• Published: 11/22/2007
• Est. Priority Date: 05/19/2006
• Status: Abandoned Application

• Current Assignee: Hitachi High-Technologies Corporation (Hitachi, Ltd.)
• Original Assignee: Hitachi High-Technologies Corporation (Hitachi, Ltd.)
• Inventors: Sakurai, Toshinari, Yamashita, Yoshihiro
• Application Number:
  US11/798,806
• Publication Number:
  US 20070269829A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 2200/0631 Purification arrangements, e.g. solid phase extraction [SPE]
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0832 cylindrical, tube shaped
• CPC Class Codes: B01L 2300/0854 Double walls
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/14 Means for pressure control
• CPC Class Codes: B01L 2400/0478 pistons
• CPC Class Codes: B01L 3/502 with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00

Abstract

A nucleic acid isolation apparatus and method enables nucleic acid isolation from a nucleic-acid-containing sample quickly with high nucleic acid yield and purity. A nucleic acid isolation instrument 11 is supported by a stand 41 via a rib 19 (A). A solution is loaded into a first container 15 via a first opening portion 14 with a loading pipette tip 42. A pressurizing/depressurizing device 43 for controlling the pressure inside the first container 15 and the second container 13 individually is closely fitted in the first opening portion 14 and the second opening portion 12 via a connecting member 44 (B). The pressurizing/depressurizing device 43 pressurizes the inside of the first container 15 while it depressurizes the inside of the second container 13 so as to move the solution from the container 15 to the container 13 via a solid phase 16 (C). After the operation of moving the solution through the solid phase 16 is conducted an appropriate number of times, the pressurizing/depressurizing device 43 is detached from the first and second opening portions 14 and 12. The solution that has moved into the second container 13 is aspirated with an unloading pipette tip 45 via the second opening portion 12 and then unloaded out of the instrument 11 (D).

Claims

1. An apparatus for isolating nucleic acid from a sample solution, comprising:

a solid phase for the adsorption of nucleic acid in the sample solution;

a nucleic acid isolation instrument comprising a first container portion and a second container portion that are communicated with each other via the solid phase wherein a solution is loaded into and unloaded out of the nucleic acid isolation instrument; and

,

pressurization/depressurization device for pressurizing the inside of one of the first container portion and the second container portion of the nucleic acid isolation instrument while depressurizing the inside of the other container portion.

2. The nucleic acid isolation apparatus according to claim 1, wherein the pressurization/depressurization device comprises a first syringe pump connected to the first container portion and a second syringe pump connected to the second container portion.

3. The nucleic acid isolation apparatus according to claim 1, wherein the sample solution is loaded into the nucleic acid isolation instrument and is unloaded out of the nucleic acid isolation instrument after a pressurization/depressurization operation by the pressurization/depressurization device, wherein a washing solution is loaded into the nucleic acid isolation instrument and the washing solution is unloaded out of the nucleic acid isolation instrument after a pressurization/depressurization operation by the pressurization/depressurization device and wherein an eluate is loaded into the nucleic acid isolation instrument and the eluate is unloaded out of the nucleic acid isolation instrument after a pressurization/depressurization operation by the pressurization/depressurization device.

4. A method of isolating nucleic acid from a sample solution, comprising:

loading a sample solution into either a first container portion or a second container portion that are communicated via a solid phase for the adsorption of nucleic acid in the sample solution;

pressurizing the inside of one of the first and the second container portions while depressurizing the other container portion so as to cause the sample solution to be reciprocated between the first container portion and the second container portion through the adsorption means and to thereby allow nucleic acid in the sample solution to become adsorbed on the solid phase; and

isolating the nucleic acid that has become adsorbed on the solid phase.

5. The nucleic acid isolation method according to claim 4, comprising:

loading, after the nucleic acid has become adsorbed on the solid phase, a washing solution into one of the first and the second container portions;

carrying out a pressurization/depressurization operation by pressurizing the inside of one of the first and the second container portions while depressurizing the inside of the other container portion so as to cause the washing solution to be moved between the first container portion and the second container portion through the solid phase;

unloading the washing solution out of a nucleic acid isolation instrument having the first container portion and the second container portion;

loading an eluate into one of the first and the second container portions;

unloading, after a pressurization/depressurization operation, the eluate out of the nucleic acid isolation instrument having the first container portion and the second container portion; and

isolating the nucleic acid.

6. An instrument for isolating nucleic acid from a sample solution, comprising:

a container;

a partition dividing the container into a first container portion having a first opening portion and a second container having a second opening portion, the partition comprising a communication opening formed therein, the communication opening communicating the first and the second container portions with each other; and

a solid phase for the adsorption of nucleic acid, the solid phase being disposed within the communication opening formed in the partition.

7. The nucleic acid isolation instrument according to claim 6, wherein the communication opening is formed at a position corresponding to the bottom of the first container portion and the second container portion.

8. The nucleic acid isolation instrument according to claim 6, wherein the bottom of the first container portion is inclined toward the bottom of the second container portion.

9. The nucleic acid isolation instrument according to claim 8, comprising a disc-shaped rib formed at the opening portion of the first container portion and the second container portion.

10. An instrument for isolating nucleic acid from a sample solution, comprising:

a container;

a partition dividing the container into a first container portion having a first opening portion and a second container portion having a second opening portion; and

a solid phase for the adsorption of nucleic acid,

wherein the first container portion comprises a communication opening formed at its bottom, the communication opening communicating the first container portion and the second container portion with each other, and wherein the solid phase is disposed within the communication opening.

11. The nucleic acid isolation instrument according to claim 10, comprising a disc-shaped rib formed at the opening portions of the first container portion and the second container portion.

12. An instrument for isolating nucleic acid from a sample solution, comprising:

a first container having an opening portion;

a second container having an opening portion; and

a solid phase for the adsorption of nucleic acid,

wherein a communication opening is formed via which the first container and the second container are communicated with each other,

wherein the solid phase is disposed within the communication opening.

13. The nucleic acid isolation instrument according to claim 12, comprising a disc-shaped rib formed at the opening portions of the first container and the second container.

14. An instrument for isolating nucleic acid from a sample solution, comprising:

a solid phase for the adsorption of nucleic acid;

a first container having a first opening portion, the first container further comprising an opening formed in its bottom in which the nucleic acid adsorption means is disposed; and

a second container having a second opening portion, wherein the second container is adapted to accommodate the first container.

15. The nucleic acid isolation instrument according to claim 14, comprising a disc-shaped rib formed at the opening portions of the first container and the second container.

16. An apparatus for isolating nucleic acid from a sample solution, comprising:

adsorption means for the adsorption of nucleic acid in the sample solution;

a nucleic acid isolation means comprising a first container portion and a second container portion that are communicated with each other via the adsorption means;

means for loading and unloading a solution into and out of the nucleic acid isolation instrument; and

pressurization/depressurization means for pressurizing the inside of one of the first container portion and the second container portion of the nucleic acid isolation means while depressurizing the inside of the other container portion.

17. The nucleic acid isolation apparatus according to claim 16, wherein the pressurization/depressurization means comprises a first syringe pump connected to the first container portion and a second syringe pump connected to the second container portion.

18. The nucleic acid isolation apparatus according to claim 16, wherein the loading/unloading means:

loads the sample solution into the nucleic acid isolation means;

unloads, after a pressurization/depressurization operation by the pressurization/depressurization means, the solution out of the nucleic acid isolation means;

loads a washing solution into the nucleic acid isolation means;

unloads, after a pressurization/depressurization operation by the pressurization/depressurization means, the washing solution out of the nucleic acid isolation means;

loads an eluate into the nucleic acid isolation means; and

unloads, after a pressurization/depressurization operation by the pressurization/depressurization means, the eluate out of the nucleic acid isolation means.