JPS62298599A - Apparatus for simultaneously performing plural reactions - Google Patents

Abstract

PURPOSE:To provide the titled apparatus enabling simultaneous synthesis of different kinds of plural DNA, etc., by stacking a plurality of flat-plate rotors each having plural circular flow channels as reaction chambers and providing the stacked rotors with rearrangeable liquid flow channels, rotating means for the rotors and liquid- feeding means. CONSTITUTION:The present apparatus 1 contains a plurality of stacked flat-plate rotors 2 each having plural circular flow channels 4, 8. Either one of the channels plays the role of reaction chamber 3 to hold a material to be reacted. A plurality of rearrangeable interconnected flow channels 4 are formed by a reaction part 6 formed by the rotors 2. These rotors 2 are independently rotated around a central axis 5 under sliding action by a rotating means. The apparatus is further provided with a group of liquid feeding lines 10 to feed different kinds of specific liquid and with a group of washing liquid feeding line 11 to feed the same washing liquid at the same time. Both line groups are connected to one of a mutually interconnected plural liquid flow channels 4 by a liquid supplying means. Different kinds of plural DNA or RNA can be synthesized at the same time by the use of a reaction apparatus 1 having the above construction.

Description

Detailed Description of the Invention 3. Detailed Description of the Invention (a) Field of Industrial Application This invention relates to a multiple simultaneous reaction device. More specifically, the present invention relates to an apparatus for synthesizing a plurality of compounds synthesized by multi-step reactions, and particularly to a multiple simultaneous reaction apparatus suitable for simultaneously synthesizing a plurality of different types of DNA or RNA (hereinafter referred to as DNA, etc.).

(B) Conventional technology Conventionally, in an apparatus for synthesizing multiple heterogeneous DNAs, etc., multiple pretreatment agents and nucleotide reagent solutions are prepared in each of multiple synthesis reaction sections each equipped with a solid support on which each nucleoside is immobilized. are introduced in a predetermined order to cause a condensation reaction,
There is a device that synthesizes by repeating this process, and in such a device, each of the above condensation reaction parts uses a liquid such as a synthetic reagent such as DNA or a liquid such as a solvent (for example, a nucleotide reagent solution, a masking agent reagent solution, a protecting group pAM reagent, etc.). A device installed in parallel with the liquid flow path of the liquid (solution, washing/drying solvent, etc.) was used.

(c) Problems to be Solved by the Invention However, in such conventional devices, each liquid flow path leading to the integrated reaction section is long, and the switching of these liquid flow paths is complicated, resulting in reaction failure due to contamination. This led to wasted reagents and equipment failure.

This invention was made in view of this situation, and is particularly applicable to processes in which the same reagent is used for multiple reactants (reagents other than nucleotide reagents, etc.), such as in the synthesis of multiple different types of DNA (reagents other than nucleotide reagents, etc.). The present invention aims to provide a multi-simultaneous reaction apparatus suitable for carrying out reactions involving a mixture of treatments using reagents (nucleotide reagents).

(d) Means for Solving the Problems Thus, according to the present invention, a flat plate is provided with a plurality of flow channels disposed circumferentially, one of which is configured as a reaction chamber capable of holding a reactant. A rotating means that configures a reaction section in which a plurality of rotors arranged in a stacked manner to set a plurality of recombinable communicating liquid flow paths, and that can independently rotate and slide these rotors around the circumferential axis. is attached to one of the plurality of communicating liquid flow paths so as to be able to switch between a group of liquid supply lines that supply different types of predetermined liquids and a group of cleaning liquid supply lines that simultaneously supply the same cleaning liquid. A multi-simultaneous reaction apparatus is provided which is characterized in that it is provided with a liquid supply means for connection.

A solid-phase reaction chamber is usually used as the reaction chamber, and is constructed by, for example, holding a carrier (resin powder or the like) on which a reactant is immobilized with a porous membrane or the like through which a liquid can pass.

Such a reaction chamber may be constructed by setting a holding area for the reactant in the flow path inside the flat rotor, but from the viewpoint of ease of attachment and detachment of carriers etc. It is preferable to protrude so that at least a flow path is formed.

Each of the flat rotors is laminated in a liquid-tight manner so as to be rotatable and slidable. At this time, in order to maintain sufficient liquid tightness, it is suitable to incorporate a backing such as an O-ring around the opening of the flow path of each rotor. Note that these plurality of flat rotors are usually held between flat stators having a liquid inlet and a liquid outlet.

As a rotating means for each flat rotor, for example, motor J5
In this case, the outer periphery of each rotor is formed in the shape of a corresponding gear.

Further, in addition to this, a shaft rotation means capable of selective rotation may be provided inside.

The plurality of liquid channels set by the stacking of the flat rotors are filled with various liquids (for example, DNA) used in multi-step reactions.
During synthesis, a 1 δT liquid supply line is connected to supply a masking agent reagent solution, a protecting group removal reagent solution, a washing drying solvent, multiple nucleotide reagent solutions, etc.; A dedicated group of cleaning liquid supply lines for supplying each liquid flow path is switchably connected to the liquid supply lines. The structure of these switching connections includes a rotating body in which through holes corresponding to a group of liquid supply lines and a group of cleaning liquid supply lines are arranged alternately around the circumference, and by rotating this body, one One example is a rotary switching valve that allows selection of a group of lines. These rotary switching valves are normally connected to each liquid flow path via a stator that sandwiches the rotor. However, in some cases, the stator itself may be provided with liquid inlet ports corresponding to a group of liquid supply lines, as well as cleaning liquid inlet ports that are in phase different from the liquid inlet ports. The liquid supply line and the cleaning liquid supply line of each group may be fixedly connected to the inlet, and any one of the groups may be selected by controlling the rotation of the rotor.

Note that the same number of cleaning liquid supply lines as each liquid flow path may be provided, but if a group of liquid supply lines includes a line for supplying cleaning liquid, it is not necessary to provide them in duplicate. Therefore, in this case, the number of cleaning liquid supply lines is one less than the number of liquid supply lines.

(E) Function According to the multiple simultaneous reaction apparatus of the present invention, each reaction chamber is connected in series and moved to a predetermined connection position with a liquid supply line by driving the rotating means, for example, in the case of processing the same reagent. By setting the settings, processing can be performed at once, and in the case of simultaneous processing of different reagents, simultaneous processing can be performed by setting each reaction chamber to a connection position with a predetermined different liquid (for example, different types of nucleotide reagent) supply line. processing becomes possible.

In addition to these different types of liquid supply lines, a dedicated cleaning liquid supply line is connected, so each liquid flow path can be cleaned at once for each step in multi-step reactions and treatments, or for each rotor rotation. , contamination between each liquid flow path is also prevented.

(f) Example FIG. 1 and FIG. 2 each show the DNA of an example of this invention.
1 is a top view and a vertical cross-sectional view showing a multiple simultaneous synthesis device 1. FIG.

Moreover, FIG. 3 is a configuration explanatory diagram that expands and explains each liquid flow path configuration of this synthesis apparatus 1. In the figure, a synthesis apparatus 1 has a disc-shaped rotor 2 configured in a reaction chamber 3 filled with a plurality of circumferential flow channels 4, one of which is filled and held with resin powder on which nucleosides are fixed. It has a plurality of reaction sections 6 arranged in a stacked manner, and these rotors can each independently rotate and slide in a liquid-tight manner around a rotating shaft 5, thereby creating a plurality of recombinable communicating liquid flow paths (flow paths). 4 and a collection of reaction chambers 3). Note that the rotating means for each rotor 2 is comprised of a selective rotating means provided on a rotating shaft and having engaging pieces that can be engaged with and released from each rotor.

As shown in FIG. 6, these selective rotation means include external rotation means consisting of a cap gear 5a provided on the outer periphery of the central portion of each rotor 2 and a motor 5C that drives the corresponding cap gear 5b. is another suitable example. In the figure, reference numeral 3a is a removable reaction chamber that is set to protrude from the side periphery of the rotor in the form of a bean gear.

Reference numerals 13 and 14 indicate disk-shaped stators that sandwich the rotor, and a liquid inlet and a liquid outlet corresponding to each liquid flow path are provided, and 8 indicates a discharge flow path.

On the other hand, below the stator 13, a flow path selection valve (7: liquid supply means) is provided which is connected to the communicating liquid flow path via the introduction path 9. The flow path selection valve 7 is a rotary valve that selectively connects each liquid supply in 1o of the group except the reaction chamber dedicated cleaning line 10' and the group of cleaning liquid supply lines 11 by rotation, and 7a is a fixed plate thereof;
7b is a rotating plate. The rotating plate 7b is provided with a plurality of liquid supply lines 10 corresponding to the liquid flow paths of the rotor, respectively.
A group of liquids for DNA synthesis, namely deprotecting group reagents 10b
, a masking reagent 10c1, an adenine-type nucleotide reagent N10d, a guanine-type nucleotide reagent M10e, a thymine-type nucleotide reagent 10r, and a cytosine-type nucleotide reagent 10g are connected, and a washing liquid 14 is connected via a plurality of branching washing liquid supply channels 11. All of these can be pumped using nitrogen gas.

The operation of such a synthesis device will be explained below.

First, the reaction chambers 3 are connected in series, and a predetermined solution is supplied by connecting these reaction chambers to a predetermined liquid inlet, and pretreatment with the same reagent is performed all at once. Next, each reaction chamber 3 is arranged so as to correspond to the intended inlet of the nucleotide reagent, respectively, as shown in FIG.
(+ is introduced into each nucleotide, and simultaneous parallel condensation reactions of nucleotides are carried out.

After the condensation reaction, each nucleotide reagent remains attached to the inner wall surface of each liquid flow path corresponding to the reaction chamber, and when the reaction chambers 3 are connected again by the rotation of the O-tor for batch post-processing, the As shown in Figure B, each residual reagent 4a is dispersed in the liquid flow path, causing contamination during the next step of simultaneous condensation reaction of nucleotides.

Therefore, before the next step of the condensation reaction, the rotary plate 7b of the flow path selection valve 7 is rotated to switch the liquid supply line 10 to the cleaning liquid supply line 11 and simultaneously introduce the cleaning liquid 14 into the liquid flow path. As a result, the remaining reagents 4a in FIG. 5B are removed to the outside of the reaction chamber, and the liquid flow paths are initialized.

By repeating the above operations, multiple simultaneous syntheses of different types of DNA can be efficiently performed.

FIG. 4 shows another embodiment in which the cleaning liquid supply line 11 is directly connected to the stator 13, and the cleaning liquid supply line 11 and the liquid supply line 10 can be selected by rotating the rotor 2. It is.

(g) Effect of the invention? What is the effect of this invention? ! According to the 2 yen 1 simultaneous reaction device, a plurality of multi-step reactions including treatments using the same reagent and treatments using different types of reagents can be easily performed at the same time. and,
In particular, contamination between these reagents can be sufficiently prevented by a dedicated cleaning liquid supply line. Therefore, it is particularly useful as a reaction apparatus for synthesizing DNA, etc., where treatments using the same reagent or different types of reagents are mixed and contamination is a problem.

[Brief explanation of the drawing]

1 and 2 are a top view and a vertical sectional view showing a multiple simultaneous DNA synthesis apparatus according to an embodiment of the present invention, and FIG. 3 is a flow path inside the synthesis apparatus of FIGS. 1 and 2. FIG. 4 is a diagram corresponding to FIG. 3 showing another embodiment; FIGS. 5A and B are explanatory diagrams showing the operation of the synthesizer of the embodiment; FIG. The figure is an explanatory diagram of the main part configuration in yet another embodiment. 1... DNA multiple simultaneous synthesis device, 2... Disc-shaped rotor, 3.3a... Reaction chamber, 4...
・Flow path, 5... Rotating shaft, 5a, 5b...
...Shade gear, 5C...Motor, 6...
...Reaction section, 7...Flow path selection valve, 7a
...Fixed plate, 7b... Rotating plate, 8.
...Discharge channel, 9...Introduction channel, 10.
...Liquid supply line, 10'...Reaction chamber dedicated cleaning line, 11...
...Cleaning liquid supply line, 12.13...Stator. Figure 1 Figure 3 Figure 4

Claims (1)

[Claims] 1. Recombination by stacking and arranging a plurality of flat rotors each having a plurality of circumferential flow channels, one of which is configured as a reaction chamber capable of holding a reactant. It constitutes a reaction section that sets a plurality of possible communicating liquid flow paths, and is provided with rotation means that can independently rotate and slide these rotors around the above-mentioned circumferential axis. A liquid supply means is provided which connects to one of the plurality of communicating liquid flow paths so as to be able to switch between a group of liquid supply lines that supply the same liquid and a group of cleaning liquid supply lines that simultaneously supply the same cleaning liquid. Features: Multiple simultaneous reaction equipment.