JPH1133436A - Angle rotor for centrifuge - Google Patents

Abstract

(57) [Problem] To provide an angle rotor which is efficiently centrifuged using a large number of test tubes and has excellent operability. A plurality of planes having an arbitrary angle with respect to a rotation axis of a rotor are provided, and a plurality of test tube insertion holes having an arbitrary angle with respect to each plane are provided on the same line. .

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an angle rotor for a centrifuge used in the fields of medicine, chemistry, pharmacy and the like.

[0002]

2. Description of the Related Art DNA sequencing processes performed in research fields such as genomic analysis, confirmation of cloned DNA, and diagnosis of pathogenic genes in the field of genetic engineering include:
DNA is separated and purified from blood cells and cells, the target gene is replicated and amplified by a method called PCR method, and then the excess fluorescent material is removed and unreacted primers are removed as the amplified product, and the DNA is taken out. The analysis has been carried out using the following device. In this step, the DNA obtained by replicating and amplifying the target gene by the PCR method is subjected to an ethanol precipitation treatment in which an appropriate amount of ethanol or the like is added and centrifuged to recover the DNA.

Conventionally, the target gene replication amplification step by the PCR method uses an apparatus called a thermal cycler.
As a sample container, 12 to 48 plastic test tubes (microtubes) of about 2 to 0.5 ml were used at the same time. After replication amplification of the target gene in a thermal cycler, the test tube was removed, and the test tube was removed from the real cycle 57-401.
Using a rotor shown in Japanese Patent Publication No. 12 (eg, the rotor shown in FIG. 4), purification of an amplification product was performed at 12,000 rpm.
The sample is centrifuged at about m (about 10,000 × g) for about 10 minutes to remove the excess fluorescent substance in the sample and the unreacted primer to remove the DNA. FIG. 4 is a vertical sectional side view showing a conventional rotor.
And a 0.2 ml micro test tube 6B is inserted on the right side through a test tube adapter. In this method, the test tube simultaneous throughput of the thermal cycler is small because the size of the test tube is limited.
It was about a book.

As an improvement of this type, recently, a capacity of 0.2
A multi-sample type thermal cycler that can use a square sample container having 96 sample injection holes called micro test tubes or micro plates each having a capacity of about 0.2 ml has been started to be sold. 0.2 ml micro test tubes are used for thermal cyclers at regular intervals.
Books and a series of 12 books are on sale. FIG. 3 shows an external view of these test tubes. In FIG. 3, (A) shows a conventionally used 1.5 ml test tube 6A, and a cap 8 connected to the test tube main body 7 by a thin member is attached to the upper end. FIG. 3B shows a 0.2 ml micro test tube 6B, and a cap 8 connected to a test tube main body 7 by a thin member is attached to the upper end similarly to the 1.5 ml test tube 6A. FIG. 3C shows a micro test tube having a structure in which twelve 0.2 ml micro test tubes are connected. Here, although the shape is slightly different from the test tube of FIG. 3B, the shape of the lower part is the same. Further, there are a series of test tubes connected by the test tube main body 7 and a series of test tubes connected by the cap 8 as shown in this figure, but there is no difference in performance and operability. In addition, adjacent intervals of the test tube main body 7 have the same dimensions, and each test tube is manufactured by molding a lot of plastic with polypropylene, and the lower portion has a substantially inverted conical shape as shown in the figure. . In addition, the interval between the test tubes having a structure in which eight or twelve 0.2 ml micro test tubes are connected is fixed to a certain value, which is the interval between the test tubes in the accommodation portion of the test tubes of the thermal cycler. Matches.

[0005]

In the conventional ethanol precipitation treatment using an angle rotor, the operation is very complicated because each test tube is handled one by one. Also,
Usually, the test tube is taken out by pinching the cap portion with a finger, but the adjacent caps hinder the removal of the test tube. In order to solve this, it is necessary to increase the interval between the adjacent test tube insertion holes, and as a result, it was not possible to provide many test tube insertion holes. Moreover, in order to mount a 0.2 ml micro test tube on a conventional rotor, it is necessary to provide an adapter for the test tube outer diameter and the test tube insertion hole diameter, which is troublesome. Further, in the 8-row or 12-row type of 0.2-ml micro test tubes, the conventional method of arranging the test-tube insertion holes of the rotor is not straight but has holes in the conical surface.
Since it had a problem that a test tube of 12 or 12 tubes could not be inserted, it could not be applied to the latest thermal cycler. In addition, the test tube for the thermal cycler has a very thin wall thickness to accurately control the temperature in the thermal cycler, and the test tube insertion hole supports the entire outer diameter of the test tube during centrifugation. If the shape is not configured, the test tube may be damaged by the liquid pressure in the test tube during centrifugation.

SUMMARY OF THE INVENTION An object of the present invention is to solve the above problems and to provide an angle rotor which can be efficiently centrifuged using a large number of test tubes and has excellent operability.

[0007]

In order to achieve the above object, a plurality of planes having an arbitrary angle with respect to the rotation axis are provided, and a plurality of test tube insertion holes having an arbitrary angle with respect to each plane are provided. This is achieved by providing them on the same line. Further, it is desirable that a plurality of test tube insertion holes are arranged at least eight at equal intervals on a straight line. Here, the reason why the number is eight or more is that a 0.2-ml micro test tube having 8 or 12 continuous test tubes is generally used.

Further, the angle of a plurality of planes on which the sample test tube insertion holes are arranged is set to 20 ° to 70 °, and the interval between the test tube insertion holes provided on the plane is 8.95 m.
m ± 0.1 mm, and is achieved by forming the lower part of the test tube insertion hole into a substantially inverted conical shape so as to match the external shape of the test tube.

[0009]

FIG. 1 is a longitudinal sectional side view showing an angle rotor according to the present invention, and FIG. 2 is a plan view showing an angle rotor according to the present invention. In FIG. 1, a rotor 1 is provided with a drive shaft hole 3 at the center thereof which is fitted and fastened to a drive shaft of a centrifuge. The plane 4, which is arranged at an angle of α ° with respect to the rotation axis 5 of the drive shaft,
The surface is provided. Further, 12 test tube insertion holes 2 are arranged at an angle of β ° with respect to each plane 4.
As a result, a total of 48 test tubes can be inserted, so that the processing efficiency can be improved. 0.2 ml on the left side of the figure
The micro test tube 6C is inserted, and twelve test tubes can be simultaneously attached and detached by picking the cap. The lower portion of the test tube insertion hole 2 has a substantially inverted conical shape as shown in the figure. Further, the rotor 1 is made of an aluminum alloy, and is designed so that the allowable maximum number of revolutions is 12,000 revolutions / minute and has sufficient durability.

In the rotor 1 configured as described above,
Each of the planes 4 provided with the test tube insertion holes 2 is at an equal distance from the rotating shaft 5 and can accommodate the same number of test tubes. α °
20 to 70 ° is appropriate, and 45 ° is most appropriate from the state of separation by centrifugation and the state of processing. The angle β ° between the test tube insertion hole 2 and the plane 5 is most preferably 90 °, and the workability is the easiest and the calmness when the test tube is inserted is good. In addition, each plane 4
The interval between the test tube insertion holes 2 provided in the sample is 8.95 mm ± 0.1
mm, which is the latest 0.2 m
The test tube interval is the same as the test tube interval of the test tube accommodating part of the thermal cycler which can use the 1 test tube. The inverted conical shape at the lower portion of the test tube insertion hole 2 substantially matches the outer shape of the 0.2 ml micro test tube described in FIGS. 3B and 3C, and the test tube is inserted. It is desirable to make the gap between the hole and the test tube very small. This can prevent the test tube from being damaged by the liquid pressure in the test tube even when the test tube with extremely thin wall pressure is rotated at a high speed and centrifuged. In addition, since the strength of the rotor 1 was set to 8,000 rpm or more, ethanol precipitation for recovering DNA was performed using a suspension obtained by adding an appropriate amount of alcohol to a solution containing DNA (nucleic acid) or protein as a liquid sample. 0.2 processing
It can be performed without any trouble by using an 8-tube or 12-tube tube of ml. In addition, since the outer diameter and the length of the test tube are small as a multiple treatment,
By providing a plurality of double holes in two or three rows, it is possible to improve the processing efficiency.

Next, using a swing rotor for a centrifuge shown in FIG.
When a rotation test was performed on each of the 8-tube and 12-tube test tubes of 2 ml micro test tubes, 12,000 rpm, 1
It was confirmed that rotation was possible without any problem up to 4,000 × g. In addition, regarding the actual centrifugation effect, a lambda DNA solution (32 g / ml), which is lambda phage DNA, was used.
An experiment for recovering DNA was carried out by using an ethanol precipitation method. When the rotation speed is 8,000 rpm (maximum centrifugal acceleration 6.3
100 × g) for 30 minutes and 12,000 rpm for 10 minutes at 12,000 × g (maximum centrifugal acceleration 14,000 × g), the DNA recovery was 100%, and the DNA recovery rate was 12,000 rpm in the conventional DNA sequencing process. The same centrifugal effect as the centrifugation could be obtained.

[0012]

According to the present invention, it is possible to provide an angle rotor which is efficiently centrifuged using a large number of test tubes and has excellent operability.

[Brief description of the drawings]

FIG. 1 is a longitudinal sectional side view showing an angle rotor according to the present invention.

FIG. 2 is a plan view showing an angle rotor according to the present invention.

FIG. 3 is an external view showing a micro test tube according to the present invention.

FIG. 4 is a vertical sectional side view showing a conventional angle rotor.

[Explanation of symbols]

1 is a rotor, 2 is a test tube insertion hole, 3 is a drive shaft hole, 4 is a plane, 5 is a rotating shaft, 6A is a 1.5 ml test tube, and 6B is 0.
A 2 ml micro test tube, 6C is a 12-tube micro test tube, 7 is a test tube main body, 8 is a cap, and 9 is a test tube adapter.

Claims (5)

[Claims] 1. An angle rotor for a centrifuge having an arbitrary angle with respect to a rotation axis of a rotor and having a test tube insertion hole capable of holding a large number of test tubes, wherein the angle rotor with respect to the rotation axis is provided. To provide a plurality of planes having an arbitrary angle,
An angle rotor for a centrifuge, wherein a plurality of test tube insertion holes having an arbitrary angle with respect to each plane are provided on the same line. 2. The angle rotor for a centrifuge according to claim 1, wherein the angle of the plane is 20 ° to 70 °. 3. The interval between the test tube insertion holes is 8.95 m.
2. The angle rotor for a centrifuge according to claim 1, wherein m ± 0.1 mm. 4. The angle rotor for a centrifuge according to claim 1, wherein a lower portion of said test tube insertion hole is formed in an inverted conical shape. 5. The angle rotor for a centrifuge according to claim 1, wherein the rotation speed of the rotor is at least 8,000 revolutions / minute.