Apparatus for purifying nucleic acids and proteins

• US 20030006193A1
• Filed: 09/06/2002
• Published: 01/09/2003
• Est. Priority Date: 09/06/1999
• Status: Active Grant

• Current Assignee: Katsunori Ikeda, Masahiro Kusumoto, Masahiro Suzuki, Yoshiaki Nishiya, Yoshihisa Kawamura
• Original Assignee: Katsunori Ikeda, Masahiro Kusumoto, Masahiro Suzuki, Yoshiaki Nishiya, Yoshihisa Kawamura
• Inventors: Suzuki, Masahiro, Nishiya, Yoshiaki, Kawamura, Yoshihisa, Kusumoto, Masahiro, Ikeda, Katsunori
• Granted Patent: US 6,986,848 B2
• Time in Patent Office: Days
• US Class Current: 210/695
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• 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 2563/143 Magnetism, e.g. magnetic label
• CPC Class Codes: G01N 1/34 Purifying; Cleaning processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003
• 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: G01N 35/028 having reaction cells in the form of microtitration plates

Abstract

The present invention provides an apparatus for purifying nucleic acids such as DNA and RNA or proteins such as enzymes and antibodies from microorganisms such as viruses and bacteria or animal and plant tissues, the apparatus being capable of fast processing of a plurality of samples. More specifically, the present invention provides an apparatus for purifying nucleic acids or proteins using magnetically attractable particles, the apparatus comprising a plurality of piston pumps; a plurality of nozzles capable of having a plurality of disposable tips which are automatically attachable/detachable; and a mechanism which is capable of dispensing a desired amount of a reagent to be used subsequently in the next step into the same number of sections as that of the samples with high accuracy and at a high rate, while a mixture of a sample and a reagent are being mixed (stirred) by the pumps and nozzles, the apparatus being capable of rapidly processing a plurality of samples, wherein a series of steps from mixing (stirring) of the magnetically attractable particles and concentration of the sample to purification is automated with reduced wastes.

Claims

1. An apparatus for purifying nucleic acids or proteins comprising:

a plurality of piston pumps; and

a plurality of nozzles capable of having disposable tips which are automatically attachable/detachable,

the apparatus being characterized by comprising;

a mechanism for mixing a liquid mixture of each sample and a reagent in a plurality of sections of a container by sucking up a plurality of the mixtures into tips, followed by discharging the mixtures in the sections simultaneously; and

a mechanism for dispensing a desired amount of a reagent to be used subsequently into the same number of sections of a different container as samples, while the mixing is in progress.

7. A method for purifying nucleic acids or proteins using the apparatus for purifying nucleic acids or proteins according to any of claims 1 to 6,

comprising sucking up each liquid not containing the objective substance into tips using a plurality of piston pumps and a plurality of nozzles capable of having disposable tips which are automatically attachable/detachable, and discharging the liquids from the tips into sections of a liquid waste container, the sections of which bottoms are inclined in a vertical cross-sectional view and being provided with holes of which diameter is smaller than the outer diameter of the tip end near the center of the bottom.

8. The method for purifying nucleic acids or proteins according to claim 7, wherein magnetically attractable particles are used.

9. The method for purifying nucleic acids or proteins according to claim 8, wherein the suspension of the magnetically attractable particles and a liquid are mixed by decreasing the pressure inside the container containing said suspension and discharging a certain amount of said suspension with the application of pressure greater than atmospheric pressure.

10. The apparatus for purifying nucleic acids or proteins according to any of claims 1 to 6, the apparatus comprising a mechanism which is capable of conducting the method of claim 9 using an air pump and an electromagnetic valve.

11. A method for purifying nucleic acids or proteins using the apparatus for purifying nucleic acids or proteins according to any of claims 1 to 6, wherein a plurality of roller tube pumps are used for transferring and dispensing the reagents, each roller tube pump being connected to tubes having different diameters, and a reagent inlet and reagent outlet of each tube being common.

12. A method for purifying nucleic acids or proteins according to claim 11, wherein dispensing is carried out alternately with normal rotation and reverse rotation of rollers.

13. The apparatus for purifying nucleic acids or proteins according to claims 1 to 6, the apparatus comprising a mechanism which is capable of conducting the method according to claim 11 or 12.

15. The apparatus for purifying nucleic acids or proteins according to any of claims 1 to 6, wherein the apparatus comprise a software which is capable of conducting the method of claim 14.

2. An apparatus for purifying nucleic acids or proteins comprising:

96 piston pumps; and

96 nozzles capable of having 96 disposable tips which are automatically attachable/detachable

the apparatus being characterized by comprising;

a mechanism for mixing a liquid mixture of each sample and a reagent in a multiple of 96 of sections of a container by sucking up a plurality of the mixtures into tips, followed by discharging the mixtures in the sections simultaneously; and

a mechanism for dispensing a desired amount of a reagent to be used subsequently into the same number of sections of a different container having sections of a multiple of 96 as samples, while the mixing is in progress.

3. An apparatus for purifying nucleic acids or proteins, the apparatus comprising:

a mechanism for removing each supernatant from a suspension of magnetically attractable particles in a multiple of 96 of sections of a container using 96 hollow cylindrical magnets; and

a mechanism for heating or cooling the container through the magnets,

wherein the purification of nucleic acids or proteins uses magnetically attractable particles.

4. An apparatus for purifying nucleic acids or proteins, the apparatus comprising:

a drive mechanism for a plurality of piston pumps;

a mechanism for driving said the drive mechanism up and down; and

magnets, a heating device and a cooling device in the direction the drive mechanism is driven.

5. An apparatus for purifying nucleic acids or proteins, the apparatus comprising:

a drive mechanism for a plurality of piston pumps;

a mechanism for driving said drive mechanism up and down;

a plate comprising a plurality of magnets having the similar arrangement to that of piston pumps directly beneath said piston pumps;

a mechanism capable of heating and cooling under said plate; and

a mechanism for conveying a container containing samples between said piston pumps and said plate.

6. An apparatus for purifying nucleic acids or proteins comprising:

a plurality of piston pumps; and

a plurality of nozzles capable of having disposable tips which are automatically attachable/detachable,

the apparatus being characterized by comprising;

a mechanism for sucking up each liquid not containing the objective substance in a plurality of sections of a container and discharging the liquids in sections of a liquid waste container, the sections of the liquid waste container having the same arrangement according to the format of the plurality of piston pumps, using a plurality of piston pumps and a plurality of nozzles capable of having disposable tips which are automatically attachable/detachable.

14. A method for purifying nucleic acids using magnetically attractable particles, the method comprising the steps of:

adding magnetically attractable particles to a cell culture solution or suspension to collect the cells;

adding a reagent for dissolving the cells to make contaminants other than an objective nucleic acid adsorb onto said particles;

separating a supernatant from said particles; and

adding another magnetically attractable particles to the supernatant.