JP2001000183A - Apparatus for preparation dna chip - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an apparatus which allows rapid and simple mass production of DNA chips by providing a means for PCR amplifying DNAs on a substrate and a means for transcription to another substrate in which mutual position of each cell of DNA for amplification is held. SOLUTION: An apparatus which allows rapid and simple mass production of DNA chips is obtained by providing a temperature-controlled incubator 10 whose inner temperature can be controlled, in which a stage 11 is provided, on which a substrate 1 is mounted, wherein plural cells and partitions for separating the cells are provided on the substrate 1 as a means for PCR amplifying DNAs which were previously prepared, by providing a means for pressing a copying substrate 2 held in holding part 13 to the substrate 1 by a driving part 14 as a means for transcripting the amplified DNA after amplification to the substrate 2 by the direct contact while keeping the mutual positions of cells.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus suitable for mass production of DNA chips.

[0002]

^2. Description of the Related Art DNA chips are generally 1 to 10 cm ^2.
, And several thousand to several hundred thousand kinds of DNAs are arranged in this region. As a method for producing a DNA chip, c prepared by a chain reaction (PCR) of a polymerase or the like was used.
A method of attaching DNA fragments to a substrate such as a slide glass or silicon using pins of an arrayer and a method of synthesizing a large number of DNAs simultaneously on a glass substrate by applying semiconductor technology are well known.

[0003]

However, there is a problem that it takes a long time to manufacture the method of attaching by means of a pin, and a large-scale factory is required for the method of applying semiconductor technology.

[0004] An object of the present invention is to solve the above-mentioned problems and to realize a DNA chip manufacturing apparatus capable of easily mass-producing DNA chips in a short time.

[0005]

In order to achieve the above object, according to the first aspect of the present invention, in a device for producing a DNA chip, a plurality of DNAs prepared in advance on a substrate serving as a matrix are subjected to a PCR method. And means for transferring the amplified DNA to another substrate by direct contact while maintaining the mutual position of each cell.

As described above, a large number of DNA chips can be manufactured easily and in a short time by the transfer method. When the DNA on the substrate serving as the mother is reduced by the transcription, the DNA can be easily amplified by the PCR method without replacing the substrate.

In this case, the amplification by the PCR method is performed on a transfer substrate which is a primary copy of the mother substrate, or in a combined state of the mother substrate and the transfer substrate. You can do so.

[0008] In another aspect of the present invention, each of the cells of the base substrate or another substrate on which DNA is transferred from the base substrate may be formed of a porous base material. it can.

According to a fourth aspect of the present invention, in the transferring means, the DNA may be transferred to another substrate by applying pressure or heating from the back of the substrate to a cell on the substrate serving as the base. . This promotes DNA transcription.

Further, a partition for separating cells may be provided between cells only when the PCR is performed. This results in perfect separation of adjacent DNA.

Further, the base substrate is formed in a cylindrical shape, and the transferring means is configured to rotate the cylindrical substrate to transfer DNA to another substrate. You can also. As a result, the area of contact at one time can be reduced, so that the restriction on the flatness is relaxed and the pressure can be easily applied.

[0012]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail with reference to the drawings. In the present invention, DNA is based on the following principle.
Mass production of chips. As shown in FIG. 1 (a), DNA is amplified by PCR on a DNA chip 1 serving as a matrix, and this is transferred to another substrate 2 while maintaining the mutual position of each cell as shown in FIG. 1 (b). Transfer by direct contact.

Next, DNA amplification and transfer are similarly performed using the substrate 2 as a mother. This is repeated to produce a large number of copies in a mouse as shown in FIG. Or,
DNA amplification from one mother substrate (DNA chip 1)
Transcription can also be performed to produce large amounts of duplicates. The PCR may be performed in a state where the mother substrate and the transfer substrate are combined.

A specific example will be described below. The cells on the mother substrate 1 are transferred substrates (called copy substrates) 2
It is not the same size as the above but a large capacity, as shown in FIG.
A plurality of porous cells (for example, sponges) 3 are aligned, and DNA is contained therein. By controlling the force pressing the copy substrate, it is possible to control the amount of image transferred in one transfer. The DNA on the mother substrate side reduced by the transcription is amplified by PCR to make up for it.

Further, as shown in FIG. 3, the amount of transcription may be controlled by extruding DNA by applying pressure or heating from the back of the chip.

The mother substrate 1 may be cylindrical as shown in FIG. 4 and transferred to the copy substrate 2 by rotation.

Furthermore, when performing PCR on the mother substrate, FIG.
As shown in (1), a cell separation partition 4 may be formed between cells (DNA spots). This septum is mechanically removed,
Alternatively, it is desirable that the substrate be formed of a substance that can be chemically removed using light, gas, or the like, and then removed after PCR.

The following transfer may be used. That is, as shown in FIG. 6, first, a solution for PCR is stored in a master substrate (each cell is made of a porous substrate), and is attached to the mother substrate by contact. The DNA attached to the mother substrate is amplified by PCR and transferred to a copy substrate, for example, in the manner shown in FIG. In addition, each cell of the mother substrate may be made of a porous substrate, a solution for PCR may be stored in the cell, and may be attached to the copy substrate by a contact.

FIG. 7 is a block diagram showing one embodiment of a DNA chip mass production apparatus to which the above-described principle of the present invention is applied. In the drawing, reference numeral 10 denotes a constant temperature bath capable of controlling the internal temperature, 11 denotes a table on which the mother substrate 1 is placed, 12 denotes a frame attached to the table 11, 13 denotes a holding unit that holds the copy substrate 2, and 14 denotes a holding unit. This is a drive mechanism that can drive the unit 13 to move it up and down. The drive mechanism 14 is attached to the frame 12.

In such a configuration, the mother board 1 of the DNA chip is placed at a predetermined position on the table 11 and the drive mechanism 14 is operated to move the copy board 2 held by the holding section 13 to the mother board 1. And press it tightly. The drive mechanism 14 can control this pressing pressure.

After the transfer to the copy board 2, the driving mechanism 14
Is operated to move the holding portion 13 upward, to separate the copy board 2 from the mother board 1 and remove it. The mother substrate 1 is left as it is. The DNA on the mother substrate side, which has been reduced by the multiple transfers, is amplified and compensated for by a PCR reaction by raising and lowering the temperature of the thermostat 10. After amplification, the temperature is lowered to a predetermined temperature.

By repeating such an operation, DN
The A chip can be easily mass-produced.

It should be noted that the foregoing description has been directed to specific preferred embodiments for the purpose of describing and illustrating the invention. Therefore, the present invention is not limited to the above-described embodiments, and includes many more modifications without departing from the spirit thereof.
This includes deformation.

[0024]

As described above, according to the present invention, there is an effect that DNA chips can be easily mass-produced in a short time.

[Brief description of the drawings]

FIG. 1 is a diagram illustrating the principle of the present invention.

FIG. 2 is a diagram illustrating another principle of the present invention.

FIG. 3 is a diagram illustrating still another principle of the present invention.

FIG. 4 is a diagram illustrating still another principle of the present invention.

FIG. 5 is a diagram illustrating still another principle of the present invention.

FIG. 6 is a diagram illustrating still another principle of the present invention.

FIG. 7 is a configuration diagram showing one embodiment of a DNA chip producing apparatus of the present invention.

[Explanation of symbols]

 REFERENCE SIGNS LIST 1 DNA chip 2 substrate 3 cell 4 cell separation partition 10 constant temperature bath 11 units 12 frame 13 holding unit 14 drive mechanism unit

Claims (6)

[Claims] 1. An apparatus for producing a DNA chip, comprising: a plurality of DNAs prepared in advance on a mother substrate;
And a means for transferring the amplified DNA to another substrate by direct contact while maintaining the mutual position of each cell. 2. The means for amplifying by the PCR method performs PCR on a transfer substrate, which is a primary copy of the mother substrate, or in a combined state of the mother substrate and the transfer substrate. The DNA chip manufacturing apparatus according to claim 1, wherein: 3. The DNA according to claim 1, wherein each cell of the base substrate or another substrate on which DNA is transferred from the base substrate is formed of a porous substrate.
Chip manufacturing equipment. 4. The method according to claim 1, wherein said transferring means is configured to promote transfer of DNA to another substrate by applying pressure or heating from the back of the substrate to a cell on the substrate serving as the base. Item 2. A DNA chip producing apparatus according to Item 1. 5. The DNA chip producing apparatus according to claim 1, wherein a partition for cell separation is provided between each cell only when the PCR is performed. 6. The substrate to be a base is formed in a cylindrical shape,
2. The DNA chip manufacturing apparatus according to claim 1, wherein said transferring means is configured to rotate the substrate serving as a base and transfer the DNA to another substrate.