KR200354278Y1 - Apparatus for controlling well temperature of high efficiency pcr device - Google Patents

Abstract

The present invention relates to a well temperature control device of a high efficiency gene amplification apparatus, and supplies a solution of different temperature to a well in which a PCR process is performed at predetermined intervals to instantly change the temperature of the well, thereby improving the yield of the gene. At the same time, we offer well temperature controls that ensure gene structure stability.

Description

Well temperature control device of high efficiency gene amplification device {APPARATUS FOR CONTROLLING WELL TEMPERATURE OF HIGH EFFICIENCY PCR DEVICE}

The present invention relates to a well temperature control device of a high efficiency gene amplification apparatus, and in particular, by forming a plurality of tanks filled with solutions of different temperatures, and supplying the solutions of these tanks to the wells for performing the PCR process at regular intervals, The present invention relates to a well temperature control device of a high efficiency gene amplification device that effectively amplifies a gene by rapidly changing the temperature of a well.

PCR (nucleic acid polymerase chain reaction) is generally performed by repeating denaturation, annealing, and elongation processes, and these three processes are performed in a heat exchanger called a well and are performed at different temperatures. . The denaturation process is 94 ℃, the annealing process is about 50 ℃, the expansion process is known as the proper temperature 72 ℃.

In the PCR process, the temperature change and the maintenance of each process have a great influence on the yield of genes, and the study of temperature control has been conducted at various angles.

Conventionally, the temperature of a PCR process was controlled using the block system using the Peltier effect. This Peltier method uses a current flow to provide thermal energy, thereby cooling or heating the block surrounding the test tube.

However, since this Peltier method is a method of heating a block, as shown in FIG. Had a problem of decreasing the production yield.

In addition, the conventional method also generates a problem that the electromagnetic waves generated by using the Peltier device negatively affect the stability of the amplified gene structure.

Therefore, the present invention is designed to solve the above problems, and an object of the present invention is to change the temperature of the well by supplying a solution having a different temperature, and controlling the temperature of the well in which the PCR process is performed. Therefore, to improve the production yield of the gene and at the same time to provide a well temperature control device of a high efficiency gene amplification device to ensure the stability of the gene structure.

1 is a graph of temperature change of a well according to a conventional well temperature controller.

2 is a circuit diagram of a well temperature control apparatus according to an embodiment of the present invention.

3 is a graph showing a temperature change of a well according to an embodiment of the present invention.

* Description of the symbols for the main parts of the drawings *

100: first tank 200: second tank

300: third tank 110: well circulation passage

111: water tank outlet pipe 112: well introduction pipe

113: well outlet pipe 114: tank return pipe

120: control unit 121: input control valve

122: return valve 130: well

140: temperature maintenance circulation circuit 141: circulation control valve

142: circulation pipe

The present invention for achieving the above object is a plurality of tanks filled with a different temperature solution; Water temperature holding means for maintaining the water temperature of the water tank; A circulation pump for circulating the solution in the tank; A well which charges a gene and transfers an externally provided heat to the gene to amplify the gene; A water tank outlet pipe formed at one end of each water tank, a well inlet pipe communicating with the water tank outlet pipe and supplying a solution of the water tank to the well, a well outlet pipe for outflowing the solution passing through the well to the outside; A well circulation passage communicating with the well outlet tube and including a water tank return tube for returning the solution back to the water tank; An input control valve formed at the outlet port of the tank to determine whether the well of the solution filled in the tank is input; A return valve formed in the tank return pipe to determine whether or not the solution has passed through the well to return to the tank; A temperature maintenance circulation circuit for continuously circulating the solution in the bath to maintain the solution temperature in each bath; A control valve for determining whether to circulate the temperature maintaining circulation passage; It provides a well temperature control device of a high efficiency gene amplification device comprising a control unit for opening or closing each of the input control valve and the return valve in accordance with the desired temperature of the well.

Preferably, the present invention includes an air spraying device for rapidly returning the solution remaining in the circulation passage to the tank when the tank for providing the solution in the well is changed, by injecting air into the circulation passage.

According to such a configuration, the present invention opens an input control valve of a water tank corresponding to a desired temperature of a well according to a control signal of a controller, thereby providing a solution having a proper water temperature in the well, and charging a solution and a gene provided in the well. The temperature of the test tube is converted to the desired temperature by heat-exchanging the test tube.

Then, when the temperature of the test tube is changed, the input control valve of the previously provided water tank is closed, and the air injection device formed in the well circulation flow path is operated to remove the solution in the well circulation flow path. By opening the input control valve of the tank to supply a solution of different water temperature to the well, the temperature of the well is changed instantaneously, thus changing the temperature of the test tube.

Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings. In addition, this embodiment is not intended to limit the scope of the present invention, but is presented by way of example only.

2 is a circuit diagram of a well temperature control apparatus according to an embodiment of the present invention, Figure 3 is a temperature change graph of the well according to an embodiment of the present invention.

As shown in FIG. 2, the present embodiment includes three tanks 100, 200, and 300 filled with solutions having different temperatures, and the three tanks maintain a water temperature to maintain a solution temperature. Means 101, 201, 301 and circulation pumps 102, 202, 302 for circulating the bath solution are formed, respectively.

The first tank 100 is filled with a 94 ℃ solution, which is the temperature required for the metamorphic action, the second tank 200 is filled with a 50 ℃ solution, which is the temperature required for the annealing process, the third tank 300 is in the expansion process Fill with 72 ° C. solution, required temperature.

And, in order to maintain each of the water temperature, the water tank 100, 200, 300 is a water temperature sensor 103, 203, 303 for sensing the water temperature of the water tank inside the water tank, respectively Heaters 104 and 204 which are installed and maintain the water temperature by heating the water tanks 100, 200, and 300 according to the signals of the water temperature sensor 103, 203, 303. 304 is formed on one side of the tank.

In addition, circulation pumps 102, 202, and 302 for supplying the solutions of the tanks 100, 200, and 300 to the outside are formed at one side of the tank, respectively.

On the other hand, the present embodiment includes a well 130 in which a process of amplifying a gene by charging a gene, transferring an externally provided heat to the gene, and the gene is performed. The well 130 is composed of a mounting portion in which a plurality of test tubes are mounted, and a well inner tube 132 passing through the periphery of the test tube, so that heat is transferred between the solution flowing in the well inner tube 132 and the test tube 131. It is formed to be made.

The wells 130 are installed in one embodiment, but a plurality of wells may be installed and a plurality of gene amplifications may be performed at one time by connecting flow channels.

Then, the tank outlet pipes 111, 211, and 311 formed at one end of each of the tanks 100, 200, and 300, and the tank outlet pipes 111, 211, and 311. A well inlet tube 133 connected to the well 130 and connected to the well 130, a well outlet tube 134 for flowing out the solution passing through the well 130 to the outside of the well, and the well outlet tube 134. And a well circulation flow path including the water tank return pipes 114, 214, and 314, which communicate with the water tank and return the solution back to the water tanks 100, 200, and 300, respectively.

The tank outlet pipes 111, 211, and 311 have input control valves 121, 221 that determine whether wells are input to the solution filled in the tanks 100, 200, and 300. 321 is formed, and the tank return pipes 114, 214, and 314 include return valves 122, 222, and 322 that determine whether the solution returns to the tank after passing through the well. Is formed.

The control unit opens or closes each of the input control valves 121, 221, and 321 and the return valves 122, 222, and 322 according to a desired temperature of the well 130. 120 is formed. This control unit 120 can be realized by a general PC.

When the input control valves 121, 221, 321 and the return valves 122, 222, 322 are opened by the control signal of the control unit 120, the respective tanks 100, ( The solution of 200, 300 is by the water pressure of the circulation pumps 102, 202, 302, the solution of each of the tanks 100, 200, 300 is the water tank outlet pipe 111 , (211), (311) → well inlet pipe (133) → well inner pipe (132) → well outlet pipe (134) → water tank return pipe (114), (214), (314) → water tank (100), Cycles in the order of (200), (300).

When the input control valves 121, 221, 321 and the return valves 122, 222, and 322 are closed by the control signal of the controller, the circulation of the solution is stopped.

On the other hand, the air injection device 150 is installed at a predetermined position of the well circulation passage. The air spraying device 150 is composed of an air compressor and a nozzle, and the air compressed in the air compressor is formed to flow into the well circulation flow path at a high speed through the nozzle.

This air spraying device sprays the high pressure air to the circulation passage when the water tank providing the solution to the well 130 is changed, and thus the solution remaining in the circulation passage (100, 200, 300). Return as soon as possible to allow the well circulation channel to cavify.

In addition, in order to maintain the solution temperature of each of the tanks 100, 200, and 300 that do not supply a solution to each of the tanks 100, 200, and 300, the solution of the tank is continuously maintained. Temperature maintaining circulation passages 140, 240, and 340 for circulating are formed, and the temperature control circulation passages include circulation control valves 141, 241, and 341 which determine whether the solution is circulated. Is formed.

The temperature maintaining circulation circuit is a circulation pipe 124, 242, (which communicates with the tank outlet pipes 111, 211, 311 and the tank return pipes 114, 214, 314). 342, and the circulation control valves 141, 241, and 341 are installed on the circulation pipes 124, 242, and 342.

When the solution is not supplied to the well 130, the input control valves 121, 221, and 321 and the return valves 122, 222, and 322 are closed, and at the same time, the circulation pipe 124 is closed. The circulation control valves 141, 241 and 341 of 242, 342 are opened.

Accordingly, the solution of the tank is the tank (100), (200), (300) → tank outlet pipes (111), (211), (311) → circulating pipes (124), (242), (342) → water It is circulated along the paths of the inquiry return pipes 114, 214, 314 → the water tank 100, 200, 300.

Referring to the operation of this embodiment configured as described above are as follows.

First, when the test tube 131 loaded with the gene is installed in the well 130, the controller opens the first tank input control valve 121 and the return valve 122 in order to perform the denaturation process of the gene. , The circulation control valve 141 is closed. At the same time, the second and third tanks of the input control valves 221, 321 and the return valves 222, 322 are closed, and the second and third tanks of the circulation control valves 241 and 341 are open. It is in the state that became.

Accordingly, the solution of 94 ° C. in the first tank is supplied to the well 130 through the water tank outlet tube 111 and the well introduction tube 133, and the test tube 131 installed in the well 130 is heated to 94 ° C. Done.

The solution of the test tube is returned to the tank through the well outlet tube 134 and the tank return tube 114, and the solution of the first tank is circulated by repeating this process.

This circulation state is maintained for 15 seconds, after which the input control valve 141 of the first water tank 100 is closed, and at the same time, the air injection device 150 is operated to remove the solution remaining in the well circulation flow path. After returning to the one tank 100 instantaneously, the return valve 122 of the first tank is closed.

Then, the circulation control valve 141 of the first tank is opened to allow the solution to circulate through the temperature maintenance circulation passage 140.

When the solution supply of the first tank 100 is stopped, the input control valve 221 and the return valve 222 of the second tank 200 are opened and at the same time the circulation control valve 241 of the second tank 200 Closed.

In this state, the solution at 50 ° C. of the second water tank 200 is supplied to the well through the second water tank outlet pipe 211 and the well introduction pipe 133 to maintain the test tube at 50 ° C., and the state is about 3 Hold for a minute to perform the gene annealing process.

When the annealing process is completed, the input control valve 221 of the second tank is closed, and at the same time, the air injection device 150 operates to instantly return the solution remaining in the well circulation passage to the second tank 200. The return valve 222 of the second water tank 200 is closed.

Then, the circulation control valve 241 of the second water tank 200 is opened to allow the solution to circulate through the temperature maintenance circulation passage 240.

When the solution supply of the second tank 200 is stopped, the input control valve 321 and the return valve 322 of the third tank 300 are opened and the circulation control valve 341 of the third tank 300 is opened. Closed.

In this state, the solution at 72 ° C. of the third water tank 300 is supplied to the well 130 through the third water tank 300, the outlet tube 311, and the well introduction tube 133, so that the test tube 131 is 72 ° C. By maintaining this state for about 30 seconds, the gene expansion process is performed.

When the expansion process is completed, the input control valve 321 of the third tank 300 is closed, and at the same time, the air spray unit 150 operates to instantly return the solution remaining in the well circulation passage to the third tank. The return valve 322 of the third tank is closed, and the circulation control valve 341 of the third tank is opened to perform the temperature maintenance circulation action.

When the solution supply of the third tank is stopped, the input control valve 121 and the return valve 122 of the first tank are opened and the circulation control valve 141 of the first tank is closed.

This action is repeated, whereby the gene denaturation, annealing, and expansion processes are repeatedly performed.

According to the present invention, by controlling the temperature of the wells by supplying solutions of different temperatures to the wells at regular intervals, the temperature of the wells is rapidly changed without time delay.

In addition, by forming an air device in the flow passage through which the solution is circulated, the influence of the solution remaining in the flow passage is reduced, and the solution of the next stage can be rapidly introduced, thereby controlling the temperature change of the well more precisely and quickly. have.

In addition, each tank to which the solution is filled forms a circulation passage for circulating the internal solution by itself so as to evenly distribute the water temperature in the tank.

Accordingly, the present invention has the effect of stably controlling the temperature of the well and at the same time shortening the temperature change time, thereby shortening the overall gene amplification time and performing accurate temperature control to significantly improve the yield of gene production. .

Claims (5)

A plurality of baths filled with solutions having different temperatures; A circulation pump for circulating the solution in the tank; A well which charges a gene and transfers an externally provided heat to the gene to amplify the gene; A water tank outlet formed at one end of each water tank, a well inlet pipe connected to the water tank outlet and connected to the well, a well outlet pipe for outflowing the solution past the well, and a water tank communicating with the well outlet pipe. A well circulation passage including a water tank return tube for returning the solution back to the furnace; An input control valve formed at the outlet port of the tank to determine whether the well of the solution filled in the tank is input; A return valve formed in the tank return pipe to determine whether or not the solution has passed through the well to return to the tank; A temperature maintenance circulation circuit for continuously circulating the solution in the bath to maintain the solution temperature in each bath; A control valve for determining whether to circulate the temperature maintaining circulation passage; Well temperature control device of the high efficiency gene amplification device including a control unit for opening or closing each of the input control valve and the return valve in accordance with the desired temperature of the well The method of claim 1, It is formed at a predetermined position of the well circulating flow path, and when the water tank provided to the well is changed, a high efficiency including an air spraying value for injecting air into the circulating flow path to quickly return the solution remaining in the circulation flow path to the water tank Well temperature control device of gene amplification device The method of claim 2, The air injection device is a well temperature control device of a high efficiency gene amplification device, characterized in that the air compressor and nozzle for providing high pressure air The method of claim 1, The temperature maintenance circulation circuit is a well temperature control device of a high efficiency gene amplification device, characterized in that to form a circulation pipe communicating the tank outlet pipe and the tank return pipe, and to form a circulation valve for controlling the circulation pipe. The method of claim 1, A plurality of the wells are formed, the well temperature control device of the high efficiency gene amplification device, characterized in that the solution of the tank is supplied to each of the plurality of wells.