KR102577197B1 - thermal block - Google Patents

Abstract

The present invention relates to thermal blocks for carrying out multiple reactions. In the present invention, a plurality of sample wells are arranged regularly in a thermal block, and a non-sample hole smaller than the sample well is provided. Two or more non-sample holes are provided within a unit area defined by connecting the center points of four adjacent sample wells among the plurality of sample wells of the thermal block. The heat block of the present invention minimizes the heat energy required for temperature change of the heat block, prevents errors in which the reaction vessel is incorrectly mounted on the heat block, and has superior durability compared to existing heat blocks.

Description

thermal block

Cross-reference to related applications

This application claims priority from Korean Patent Application No. 10-2018-0074957, filed with the Korean Intellectual Property Office on June 28, 2018, the disclosure of which is incorporated herein by reference in its entirety.

Technology field

The present invention relates to thermal blocks for carrying out multiple reactions.

The most commonly used nucleic acid amplification reaction, known as polynucleotide chain reaction (PCR), involves repeated cycles of denaturation of double-stranded DNA, annealing of oligonucleotide primers to a DNA template, and primer extension by DNA polymerase. (Mullis et al., US Pat. Nos. 4,683,195, 4,683,202 and 4,800,159; Saiki et al., (1985) Science 230, 1350-1354).

In order to detect the same target in a plurality of samples, a medical examination center that performs tests on samples collected from many patients simultaneously amplifies the plurality of samples in parallel under predetermined reaction conditions using a reaction mixture of a predetermined composition. Therefore, PCR devices used to detect target nucleic acid sequences from samples are designed to amplify multiple samples simultaneously. For example, a PCR device includes a thermal block that can accommodate a plurality of reaction tubes. In addition, it is equipped with a device and control system that can quickly change the temperature of each well (the groove that accommodates the reaction vessel) and maintain it uniformly, and is a program that can read each result performed in multiple wells and report comprehensively. Includes. In addition, a reaction plate in the form of a plurality of reaction tubes connected that can be used in the above PCR device has been developed and used.

Thermal block, also called a heating block, has a plurality of wells to accommodate a plurality of reaction vessels and is made of metal for rapid heat conduction. However, metal has a high specific gravity and specific heat, so there is a problem in that a large amount of energy must be supplied or removed to control the temperature of the heat block.

As a method to solve this problem, US7955573, US7632464, and US8557196 disclose a method of making holes in the side and top of the thermal block. The goal is to change the temperature of the heat block more quickly by removing unnecessary parts of the heat block. However, there was a problem with users placing the reaction plate in the hole formed in the upper surface, and to solve this problem, a thermal block was developed in which the hole located in the center of the thermal block among the plurality of holes formed in the upper surface was removed.

However, this configuration does not solve the problem of incorrect installation of reaction vessels that occupy some space in the thermal block, such as a single PCR tube or 8 PCR tube strips, and furthermore, the pattern of the sample wells and mass reduction holes formed in the thermal block is not uniform. Therefore, another problem arises in which a temperature difference occurs between the reaction plate or a plurality of reaction tubes accommodated in each well.

The PCR reaction is a reaction in which the target nucleic acid sequence is amplified by hybridizing a specific primer to the target nucleic acid sequence, extending it with polymerase, and then repeating the steps of separating the extended strands. The PCR reaction efficiently performs this series of steps by maintaining the reaction mixture at specified temperatures for a specified period of time. Therefore, it is very important to maintain the correct temperature at each step in the PCR reaction. This is because the reaction efficiency at each stage may vary depending on the temperature.

In particular, when performing the same test on multiple samples using a PCR reaction, the temperature difference that continuously occurs between wells causes the amplification reaction to proceed at different efficiencies for multiple samples in which the amplification reaction was performed in different wells. do. The PCR reaction repeats the nucleic acid amplification reaction for dozens of cycles, and since the nucleic acid strand generated in the previous cycle becomes the template for the next cycle, the difference in amplification efficiency that occurs in each cycle can have a significant impact on the analysis results. there is.

Meanwhile, US7081600 discloses a thermal block in which inverted cone-shaped sample wells connected to a support at the bottom are arranged in a uniform manner. In this type of thermal block, the upper part of the sample well is all independently disconnected. In this case, the effect of mass reduction increases, but there may be problems with physical durability.

In order to preserve the high-temperature material in the tube during the PCR process and prevent the lid of the reaction tube from opening, the PCR device includes a hot lid that can be pressed against the top of the reaction tube to prevent it from opening. The hot lid is adhered to the top of the reaction tube and maintained while applying pressure to the reaction cube. The pressure is transmitted to the heat block through the reaction vessel, thereby deforming the heat block. The weaker the horizontal connection between each sample well, the weaker the durability of these sample wells. In particular, if the plate or reaction tube is installed incorrectly and the hot lead is covered, deformation or damage to the sample well of the heat block may occur.

Therefore, there is a need to develop a new thermal block that can minimize temperature differences between the reaction plate or a plurality of reaction tubes, while simultaneously preventing errors in incorrectly mounting the reaction plate, and has excellent durability.

Against this background, the purpose of the present invention is to provide a thermal block that has a uniform non-sample hole pattern and is excellent in durability without the problem of mismounting the reaction plate.

The present invention is a thermal block for performing multiple reactions,

The thermal blocks are parallel to each other and have upper and lower surfaces having a length and width,

A plurality of sample wells open to the upper surface are formed on the upper surface, and the plurality of sample wells are arranged regularly, and

Two or more non-sample holes open to the upper surface are formed on the upper surface, and the upper surface opening area of each non-sample hole is smaller than the upper surface opening area of the sample well,

The purpose is to provide a thermal block characterized in that two or more non-sample holes are included in a unit area defined by connecting the center points of four sample wells adjacent to each other in the thermal block.

To achieve the above-described object, in one aspect, the present invention

A thermal block for performing multiple reactions,

The thermal blocks are parallel to each other and have upper and lower surfaces having a length and width,

A plurality of sample wells open to the upper surface are formed on the upper surface, and the plurality of sample wells are arranged regularly, and

Two or more non-sample holes open to the upper surface are formed on the upper surface, and the upper surface opening area of each non-sample hole is smaller than the upper surface opening area of the sample well,

A thermal block is provided, wherein two or more non-sample holes are included in a unit area defined by connecting the center points of four sample wells adjacent to each other in the thermal block.

The heat block of the present invention removes unnecessary parts of the heat block, minimizing the heat energy required for temperature changes in the heat block, and preventing errors in incorrectly mounting the reaction vessel.

In the thermal block according to an embodiment of the present invention, the opening of the non-sample hole formed to reduce unnecessary mass of the thermal block has a smaller area than the opening of the sample well, thereby preventing errors in which the reaction vessel is incorrectly mounted.

Meanwhile, the thermal block of the present invention has two or more non-sample holes within a unit area, maximizing the reduction of unnecessary mass within the thermal block while using non-sample holes smaller than the sample well.

In addition, the non-sample holes are formed so that the formation patterns of two or more non-sample holes formed in each unit area within the thermal block are the same between each unit area, thereby reducing the variation between each sample well.

The non-sample hole may be formed so that the thermal conductive effect of the non-sample hole within the unit area is the same for each of the four adjacent sample wells defining the unit area, and this configuration allows each sample of the thermal block to have the same thermal conductive effect. Improves thermal uniformity between wells.

In addition, when each sample well is connected in a straight line to adjacent sample wells, including sample wells located diagonally, the area connected by the straight line serves as a support wall between each sample well, which has the effect of increasing the overall durability of the thermal block. there is.

Figure 1 is a perspective view of a thermal block of the present invention.
Figure 2 is a plan view of the thermal block of the present invention.
Figure 3 is a schematic diagram showing the arrangement of sample wells and non-sample holes formed in the thermal block of the present invention.
Figure 4 is a schematic diagram showing the arrangement of non-sample holes provided in the thermal block of the present invention. The non-sample holes are arranged so that the heat conduction influence of the non-sample holes within the unit area for each of four adjacent sample wells is the same.
Figure 5 is a cross-sectional view showing the cross-section of the sample well and the non-sample hole of the present invention taken along line AB shown in Figure 2.
Figure 6 is a cross-sectional view of the thermal block of the present invention along the CD shown in Figure 2, cut through the center point of the sample wells parallel in the width direction. It can be seen that no non-sample holes are formed.
Figure 7 is a cross-sectional view of the thermal block of the present invention cut along the plane passing through the center point of the sample well in the diagonal direction of the thermal block according to EF shown in Figure 2, and it can be seen that no non-sample hole is formed.
Figure 8 shows the results of comparing the ramp rate when using the thermal block of the present invention and the control thermal block without a non-sample hole, respectively.

Hereinafter, the present invention will be described in detail through exemplary drawings.

When adding reference numerals to components in each drawing, it should be noted that identical components are given the same reference numerals as much as possible even if they are shown in different drawings. Additionally, in describing the present invention, if it is determined that a detailed description of a related known configuration or function may obscure the gist of the present invention, the detailed description will be omitted.

Additionally, when describing the components of the present invention, terms such as first, second, A, B, (a), and (b) may be used. These terms are only used to distinguish the component from other components, and the nature, sequence, or order of the component is not limited by the term. When a component is described as being “connected,” “coupled,” or “connected” to another component, that component may be directly connected or connected to that other component, but there is another component between each component. It will be understood that elements may be “connected,” “combined,” or “connected.”

Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings in order to enable those skilled in the art to easily practice the present invention.

Figure 1 is a perspective view of a thermal block according to an embodiment of the present invention. Figure 2 is a plan view of a thermal block according to an embodiment of the present invention. Figure 6 is a cross-sectional view of a thermal block according to an embodiment of the present invention.

Referring to Figures 1 and 6, the thermal block 100 of the present invention has a top surface 101 and a bottom surface 102 having a length and a width, and the top surface 101 and the bottom surface 102 (102) are parallel to each other. The thermal block 100 of the present invention may have the shape of a hexahedron, particularly a rectangular parallelepiped, with a constant height (thickness). The length and width of the upper and lower surfaces may be different, and the front, left, and right surfaces may be curved depending on the shape of the sample hole and/or non-sample hole.

The thermal block 100 of the present invention can be used as a reaction vessel in which a plurality of wells formed in the thermal block directly receive and react with samples, or a receptor that accommodates a reaction vessel formed to fit the plurality of wells formed in the thermal block. It can be used as According to one embodiment of the present invention, the thermal block 100 may be manufactured using a material with excellent thermal conductivity. It may be made of metal or metal alloy (e.g., iron, copper, aluminum, gold, silver, or alloys containing the same). The thermal block may be machined from a single solid piece of metal, or may be formed by connecting several metal pieces.

The thermal block 100 of the present invention is a thermal block for performing multiple reactions. The reaction refers to a chemical, biochemical or biological transformation involving at least one chemical or biological substance (eg, solution, solvent, enzyme). In the present invention, the reaction may preferably be initiated, stopped, promoted, or inhibited by thermal changes in the reaction system. For example, the reaction may be a reaction in which the decomposition or combination of biological or chemical substances progresses depending on temperature changes, or the activity of enzymes that produce or decompose biological or chemical substances may be promoted or inhibited depending on temperature changes. .

Specifically, the reaction may refer to an amplification reaction. The amplification reaction may be a reaction that increases the target analyte (eg, nucleic acid molecule) itself, or may be a reaction that increases or decreases a signal generated depending on the presence of the target analyte. A reaction that increases or decreases the signal generated depending on the presence of the target analyte may or may not be accompanied by an increase in the target analyte. Specifically, the target analyte is a nucleic acid molecule, and the reaction may be polymerase chain reaction (PCR) or real-time PCR.

According to one embodiment of the present invention, the thickness of the thermal block 100 of the present invention may be 5 mm to 20 mm. If it is 5 mm or less, the sample well 103 formed in the heat block 100 may not be formed sufficiently to accommodate the reaction vessel, and if it is 20 mm or more, too much heat energy is required to change the temperature of the heat block 100. It may be inefficient because it is necessary.

The length and width of the thermal block 100 may vary depending on the size and number of sample wells 103 formed in the thermal block 100. The length and width of the thermal block 100 may each independently be 10 mm or more, 20 mm or more, or 30 mm or more. In addition, the length and width of the thermal block 100 may each independently be 1000 mm or less, 900 mm or less, 800 mm or less, 700 mm or less, 600 mm or less, 500 mm or less, 400 mm or less, 300 mm or less, or 200 mm or less.

The thermal block 100 may include a recessed portion in which a temperature sensor can be mounted. The recessed portion for mounting the temperature sensor may be formed on the top, bottom, or side of the heat block, and may specifically be formed on the bottom. The recessed portion may vary depending on the shape of the temperature sensor, and for example, may be formed in a shape in which a probe-type or button-type temperature probe can be mounted. According to one embodiment of the present invention, a bar-shaped depression may be formed on the lower surface of the thermal block of the present invention so that a probe-type temperature sensor can be mounted. In addition, the heat block 100 includes a reaction vessel in which a device for supplying optical or electrical energy to the reaction vessel and/or a device for detecting an optical or electrical signal emitted from the reaction vessel is accommodated in each sample well 103. An additional contact path may be included. For example, a receiving hole capable of accommodating a light source or light detector may be included on the bottom or side of the sample well 103. The receiving port may directly receive a light source such as an LED or a light detector such as a photodiode, or may receive an optical fiber connected to such a light source or a light detector.

According to one embodiment of the present invention, the lower surface 102 of the thermal block 100 may be formed as a plane so that a thermoelectric element such as a Peltier element can contact it.

The thermoelectric element may serve as a heating element that supplies heat when electrical energy is provided, or it may serve as a cooling element that absorbs heat. Depending on the heating and cooling of the thermoelectric element, the heat block can transfer heat to the reaction vessel contained in the sample well or absorb heat from the reaction vessel.

A plurality of sample wells 103 open to the upper surface 101 are formed on the upper surface 101 of the thermal block of the present invention. The sample well 103 of the present invention is formed on the upper surface 101 of the thermal block and can directly accommodate a sample or a reaction vessel formed to fit the sample well 103. The sample well 103 is formed in a size and shape that allows it to thermally-conductively contact the reaction vessel.

The thermal block 100 of the present invention is designed to perform multiple reactions simultaneously. The sample well 103 formed in the thermal block 100 is a plurality of sample wells. The thermal block 100 may include 4 or more, 6 or more, 8 or more, 12 or more, 16 or more, 24 or more, 32 or more, 40 or more, and 48 or more sample wells 103. The thermal block 100 may include sample wells 103 of 96 or less, 192 or less, 288 or less, and 384 or less. According to one embodiment of the present invention, the thermal block 100 may include 4 to 384 sample wells 103 open to the upper surface 101.

The plurality of sample wells 103 may be arranged regularly.

The regular arrangement means that the direction and distance of one sample well 103 among the plurality of sample wells 103 with respect to the adjacent sample well 103 are determined according to a certain rule. The arrangement of the plurality of sample wells 103 is determined by the above rules.

For example, the plurality of sample wells 103 of the present invention may be arranged side by side in two or more rows parallel to each other in the first direction on the upper surface 101, and among the plurality of sample wells 103, Sample wells 103 arranged in each row may be arranged at regular intervals.

According to one embodiment of the present invention, the regular arrangement may be a rectangular array. The rectangular array includes a plurality of sample wells 103 arranged side by side in two or more rows parallel to each other in the first direction, two or more rows parallel to each other in a second direction perpendicular to the first direction ( This means that they are arranged side by side in columns. Among the plurality of sample wells 103, the sample wells 103 arranged in each row in the second direction may be arranged at regular intervals.

According to one embodiment of the present invention, the regular arrangement may be a square arrangement. The square array is a special form of the rectangular array, and refers to an arrangement in which the spacing between the sample wells 103 arranged in the first direction is the same as the spacing between the sample wells 103 arranged in the second direction. When a plurality of sample wells 103 are arranged in a square arrangement, the distance between the sample wells 103 in each row is the same, so a reaction vessel containing a plurality of containers (for example, in one column) 8 PCR tube strips containing 8 connected containers) can be mounted on the heat block 100 without considering directionality.

According to one embodiment of the present invention, the thermal block 100 of the present invention is not limited thereto, but includes 96 sample wells 103 arranged in a grid of 12 rows and 8 columns, and a grid of 4 rows and 4 columns. It may include 16 sample wells 103 arranged, or 384 sample wells 103 arranged in a grid of 24 rows and 16 columns.

According to one embodiment of the present invention, the sample well 103 of the present invention may be formed to accommodate a reaction vessel. The reaction vessel may be a reaction tube containing one container, or may be a reaction strip or reaction plate containing a plurality of containers. The reaction strip refers to a reaction vessel in which a plurality of containers are formed in a row at regular intervals, and the reaction plate refers to a reaction vessel in which a plurality of containers are formed in two or more rows at regular intervals. The container refers to a unit that can accommodate a reactant (eg, a reaction solution or a reaction mixture). The reaction vessel may be named a test tube, PCR tube, strip tube, or multi-well PCR plate depending on its purpose and form.

The sample well 103 of the present invention may vary depending on the shape of the container of the reaction vessel used. For example, the sample well 103 of the present invention can be formed to accommodate a general conical tube for nucleic acid amplification. In this case, the sample well 103 has a circular opening opened to the upper surface 101 of the heat block, and is tapered so that the diameter of the bottom surface 105 of the sample well is narrower than that of the opening 104 opened to the upper surface 101. It can be. Figure 5 shows a cross-section of an exemplary sample well. As shown in FIG. 5, each sample well 103 may be formed to be tapered so that the outer surface of the reaction vessel can come into close contact with the inner wall (inner surface) of the sample well.

According to one embodiment of the present invention, the sample well 103 is a container having a volume that can accommodate 10 microliters or more, 20 microliters or more, 30 microliters or more, and 40 microliters or more of reaction solution for nucleic acid amplification. It can be formed to accommodate a reaction vessel containing.

According to one embodiment of the present invention, the sample well 103 is 700 microliters or less, 600 microliters or less, 500 microliters or less, 400 microliters or less, 300 microliters or less, 200 microliters or less, 100 microliters or less. Alternatively, it may be formed to accommodate a reaction vessel including a container having a volume capable of accommodating a reaction solution for nucleic acid amplification of 50 microliters or less.

The thermal block 100 includes two or more non-sample holes 106. The non-sample hole 106 is formed on the upper surface 101 of the thermal block and is formed to open to the upper surface 101. The non-sample hole 106 is separated from the sample well 103, and the non-sample hole 106 does not accommodate a reaction vessel. The non-sample hole 106 is provided to reduce the energy required to change the temperature of the sample well 103.

The shape of the non-sample hole 106 is not particularly limited, but may be formed so that a reaction vessel fitted to the sample well 103 cannot be inserted. A reaction vessel of a type suitable for the sample well 103 refers to a reaction vessel formed so that the outer surface of the reaction vessel is in close contact with the inner wall of the sample well 103 when installed. The shape of the non-sample hole 106 is not limited to this, but may be, for example, a circle, an oval, or a polygon such as a square or triangle. According to one embodiment of the present invention, the non-sample hole 106 of the present invention may have a cylindrical structure with a circular or oval upper surface.

The area of the opening 107 of the non-sample hole formed on the upper surface of the thermal block 100 is smaller than the area of the opening 104 of the sample well. Since the area of the opening 107 of the non-sample hole is smaller than the area of the opening 104 of the sample well, the risk of the user incorrectly mounting the reaction vessel to be mounted on the sample well 103 into the non-sample hole 106 can be prevented. there is. If an incorrect mounting prevention configuration is included in the center of the thermal block, such as in US 8367014, incorrect mounting of a reaction vessel that covers the entire thermal block, such as a 96 plate, can be prevented, but reaction vessels such as a single PCR tube or 8 PCR tube strips can be prevented. Improper installation of the container cannot be prevented. On the other hand, in the thermal block 100 of the present invention, the area of the opening 107 of the non-sample hole is smaller than the area of the opening 104 of the sample well, so the reaction vessel can be used regardless of the type of reaction vessel and the position at which the reaction vessel is mounted on the thermal block. This can prevent it from being installed incorrectly.

According to one embodiment of the present invention, the opening areas of the non-sample holes may be different. The non-sample holes of the present invention do not need to have the same overall shape and size. For example, additional small non-sample holes may be formed in the space between non-sample holes formed within a unit area.

The non-sample hole 106 may be formed in the same pattern as the thermal block 100. Formed in the same pattern means that when a plurality of shapes with the same shape and area are drawn by connecting the center points of the sample wells 103 in the thermal block 100, the shape and area are compared within the plurality of shapes with the same shape and area. This means that the number and distribution form of the sample holes 106 are formed to be the same.

Specifically, the non-sample hole 106 is formed so that the patterns of the non-sample holes in the unit area 130 defined by connecting the center points of four adjacent sample wells 103 in the thermal block 100 are the same. .

The four sample wells 103a to 103d adjacent to each other refer to a combination of the four sample wells closest to each other among the sample wells formed on the upper surface 101 of the thermal block. Referring to Figures 3 and 4, the combination of the four sample wells closest to each other includes one sample well (103a) and the other sample well (103b) closest to the one sample well (103a). It can be determined by additionally determining two sample wells (103c, 103d) that are not parallel to and closest to the determined two sample wells (103a, 103b). The fact that the two sample wells (103c, 103d) are not parallel to the two sample wells (103a, 103b) means that the two sample wells (103c, 103d) are a straight line connecting the center points of the two sample wells (103a, 103b). It means that it is not located on the top.

According to one embodiment of the present invention, the four adjacent sample wells 103a-103d are closest to each other and may be a combination of four sample wells that can form a square by connecting their center points.

The unit area 130 is an area defined to express the pattern of the sample well 103 and the non-sample hole 106 of the thermal block 100 of the present invention. The unit region 130 refers to a region formed by connecting the center points of the four adjacent sample wells 103a-103d.

The part of the unit area excluding the space occupied by the sample wells is defined as a mass region. The space occupied by the four adjacent sample wells is not included in the mass area. Therefore, the mass area is an area where the reaction vessel is not located. Accordingly, the smaller the mass of the mass region, the smaller the amount of heat energy change required to change the reactants in the reaction vessel to the desired temperature.

As shown in FIG. 3, according to one embodiment of the present invention, the mass area is an area in which sample wells are not formed among the areas formed by connecting the center points of the four adjacent sample wells 103a to 103d. You can. According to one embodiment of the present invention, the mass area may be an area excluding the area occupied by the sample wells in a rectangular area formed by connecting the center points of the four adjacent sample wells 103a to 103d.

The non-sample hole 106 of the thermal block 100 may be formed in the mass area. By forming the non-sample hole 106, the mass of the mass area can be reduced, and thus the amount of heat energy change required to change the reactants in the reaction vessel to the desired temperature can be reduced.

According to one embodiment of the present invention, the patterns of the non-sample holes 106 formed in the unit area 130 may be the same. This means that the distribution (relative position), size, and shape of the non-sample holes 106 formed in the plurality of unit areas 130 defined by four sample wells adjacent to each other in the thermal block 100 are the same. It means that The dotted area in FIG. 2 arbitrarily indicates unit areas 130 that can be set in the thermal block of the present invention. As shown in FIG. 2, all unit regions 130 formed by four sample wells adjacent to each other in the thermal block 100 of the present invention have the same pattern of non-sample holes 106 formed therein. .

Since the plurality of sample wells 103 are regularly arranged in the thermal block 100 of the present invention, the unit areas 130 within the thermal block 100 all have the same shape and area. Therefore, when the non-sample holes 106 in the unit area 130 are formed in the same pattern, the thermal conductive effect of the non-sample holes 106 on all sample wells 103 is the same. . This can minimize temperature differences between each sample well that may occur due to rapid heating or cooling of the thermal block. Although it is common to set the unit area 130 so that all non-sample holes 106 of a thermal block are included in one of the unit areas 130 that can be defined in the thermal block, the setting of the unit area 130 Accordingly, some non-sample holes 106 may not be included in the unit area 130. These non-sample holes 106 can be arranged considering the arrangement pattern of the non-sample holes 106 across the entire thermal block.

According to one implementation of the present invention, one non-sample hole 106 may be formed in one unit area 130. In other words, all non-sample holes 106 can be formed within the unit area 130 without the non-sample holes 106 being formed across two or more unit areas 130 . When the non-sample hole 106 is formed in this way, the area connecting the sample wells 103 is not cut off by the non-sample hole 106, and thus the durability of the thermal block 100 can be increased. Meanwhile, alternatively, according to one implementation of the present invention, one non-sample hole 106 may be formed over two or more adjacent unit areas 130. If the pattern of the internal non-sample holes 106 is the same for each unit area 130, one non-sample hole 106 can be formed over two or more adjacent unit areas 130.

According to one embodiment of the present invention, the unit area 130 may be a minimum rectangular area defined by connecting the center points of four adjacent sample wells in the thermal block 100, and the non-sample hole 106 may be formed so that the patterns of the non-sample holes 106 in all of the minimum rectangular regions are the same.

The minimum square is created by selecting four sample wells (103a to 103d) that can form a square by connecting the center points of the sample wells 103 formed on the upper surface 101 of the thermal block, and connecting their center points. Among the rectangles that can be defined, it refers to the rectangle whose area is the minimum and whose sum of the lengths of the four sides of the rectangle is the minimum.

Meanwhile, in the non-sample hole 106, the opening 107 of the non-sample hole is formed to be smaller than the opening 104 of the sample well. Therefore, in order to improve the effect of reducing the mass of the entire thermal block 100 due to the formation of the non-sample holes 106, the number of non-sample holes 106 formed in the thermal block is greater than the number of sample wells 103. It can be.

According to one embodiment of the present invention, two or more non-sample holes 106 are included in the unit area 130 defined by connecting the center points of four adjacent sample wells in the thermal block 100. Do it as

According to one embodiment of the present invention, the number of non-sample holes 106 formed in the unit area 130 defined by connecting the center points of four adjacent sample wells in the thermal block 100 is a multiple of 2. You can. When the non-sample holes 106 in the unit area 130 are formed in multiples of 2, the non-sample holes 106 in the unit area 130 are each of the four sample wells forming the unit area 130. It can be formed identically or symmetrically with respect to the center point. The number of non-sample holes 106 formed in the unit area 130 may be a multiple of 4.

According to one embodiment of the present invention, four or more non-sample holes 106 may be formed in the unit area 130 defined by connecting the center points of four sample wells adjacent to each other in the thermal block. Specifically, the number of non-sample holes 106 formed in the unit area 130 is not limited to this, but may be, for example, 4 to 100, 4 to 48, 4 to 24, or 4 to 16. . According to one embodiment of the present invention, the number of non-sample holes 106 included in the unit area is not limited to this, but is 2, 4, 6, 8, 10, 12, and 14. Or it could be 16.

According to one embodiment of the present invention, the non-sample hole 106 may be formed so that the thermal conductive influence of the non-sample hole 106 within the unit area 130 for each of the four adjacent sample wells is the same. there is.

When the non-sample hole 106 formed in the unit area 130 is formed to provide the same thermally conductive effect to each of the four adjacent sample wells defining the unit area 130, the thermal block 100 ) Each sample well 103 formed within is subject to the same heat conduction influence by the surrounding non-sample holes 106. This helps maintain thermal uniformity between each sample well. The non-sample hole 106 formed in the unit area 130 is formed to provide the same thermal conductivity effect to each of the four adjacent sample wells, for example, to the four sample wells adjacent to each other in the thermal block. The non-sample holes 106 in a unit area defined by connecting the center points of may be identical or formed symmetrically with respect to the center points of each of the four sample wells defining the unit area 130.

According to one embodiment of the present invention, the non-sample holes 106 in the unit area 130 may be formed identically or symmetrically with respect to the center point of each of the four sample wells forming the unit area 130. You can. This may indicate that the four adjacent sample wells have the same positional relationship to the non-sample holes within the unit area 130. In addition, a figure drawn by connecting a sample well of one of the four sample wells and the non-sample holes in the unit area 130 is a ratio of a sample well of another one of the four sample wells and the non-sample holes in the unit area 130. It may be identical to or symmetrical to the figure drawn by connecting the sample holes.

In other words, for each of the four sample wells forming the unit area 130, the non-sample holes 106 formed within the unit area 130 may be arranged in the same or symmetrical pattern. Figure 3 shows four sample wells (103; 103a, 103b, 103c, 103d) adjacent to each other in a thermal block, a unit area 130 defined by them, and a non-sample hole 106 within the unit area 130. It was done. FIG. 4 shows non-sample holes 106 and the four sample wells 103a, 103b, 103c, and 103d in a unit area 130 defined by connecting the center points of four adjacent sample wells shown in FIG. 3. ), each figure is drawn by connecting one of them. As shown in Figure 4, it is drawn by connecting one sample well of the four sample wells (103a, 103b, 103c, 103d) and two or more non-sample holes (106) included in the unit area (130). The shape and size of the figure are the same as those formed by connecting another sample well among the four sample wells and the non-sample hole 106 included in the unit area 130, or are the same in size. And the shape can be formed symmetrically as a mirror image.

In this way, when the non-sample holes 106 included in the unit area 130 provide the same or symmetrical pattern for each of the four sample wells forming the unit area 130, the unit area (130) The non-sample hole group may provide the same thermal effect to the sample wells. Therefore, the thermal block of the present invention can maintain thermal uniformity between each sample well.

As described above, the regular arrangement of the plurality of sample wells 103 may be a rectangular array.

According to one embodiment of the present invention, the regular arrangement of the plurality of sample wells 103 is a rectangular arrangement, and the diagonal of the unit area 130 is at least one non-sample hole 106 in the unit area 130. ) may touch or not intersect. The contact means that one non-sample hole and one straight line contact at one point, and the one point of contact is called a tangent point. The intersection means that one straight line touches one non-sample hole at two points, and the two touching points are called intersections. The diagonal line of each unit area 130 may be defined by connecting the center points of four adjacent sample wells defining each unit area 130.

According to one embodiment of the present invention, the regular arrangement of the plurality of sample wells 103 is a rectangular arrangement, and the diagonal line formed by connecting the center points of four adjacent sample wells defining the unit area 130 is It may contact or not intersect all non-sample holes 106 within the unit area 130. In other words, the non-sample hole 106 in the unit area 130, which is formed by connecting the center points of the four most adjacent sample wells, can be formed so that no non-sample hole includes the diagonal line of the unit area 130. there is. The non-sample hole 106 provided in the thermal block 100 of the present invention may be formed so that the diagonal line connecting the sample well 103 does not contact or intersect the non-sample hole 106. Therefore, in this case, As can be seen in FIG. 7, the non-sample hole 106 does not appear in a cross-sectional view of the thermal block 100 of the present invention cut diagonally (for example, along line E-F in FIG. 2).

In this way, if the non-sample hole 103 is formed not to contact or intersect the diagonal line connecting the sample well 103, each sample well 103 in the thermal block 100 is adjacent to the diagonal line among the remaining sample wells. It is physically connected to the sample well in a straight line.

In the existing sample block or thermal block, a mass reduction hole of a considerable size is formed in the center of the mass area within the unit area 130. In a thermal block of this type, each sample well is physically connected to the four adjacent sample wells in a straight line, but is not physically connected to the diagonally adjacent sample wells in a straight line. According to another known form, each sample well in an inverted cone shape is connected only through the support part at the bottom, and the upper parts are all independently disconnected. Moreover, in the case where inverse conical sample wells are formed independently as in the latter case, each sample well is physically disconnected from adjacent sample wells except for the support at the bottom. Therefore, there is a problem in that the physical durability of sample wells formed in this way is weak.

On the other hand, in the thermal block 100 of the present invention, each sample well 103 is physically connected to eight adjacent sample wells in a straight line. Therefore, the heat block 100 of the present invention reduces the amount of change in heat energy required to change the temperature of the reactants in the reaction vessel to the desired temperature by reducing the mass of the mass area, and at the same time provides superior durability compared to the existing heat block. You can.

According to one embodiment of the present invention, in the thermal block 100 of the present invention, all straight lines connecting the center points of four adjacent sample wells defining the unit area 130 are non-sample areas within the unit area 130. It may be formed so as not to contact or intersect the hole 106. In other words, the non-sample hole 106 is not formed on any straight line connecting the center points of the four sample wells forming the unit area 130. As can be seen in FIGS. 6 and 7, the thermal block 100 of the present invention has no non-sample cross section along a straight line connecting the center points of the sample wells (for example, lines C-D and lines E-F in FIG. 2). The hole 106 may also be formed not to appear. All straight lines connecting the center points of the four sample wells include four sides and two diagonals of a square formed by connecting the center points of the four sample wells.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it is understood by those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. It will be self-evident.

Example 1: Thermal Block Production

The well size of the thermal block of the present invention can be freely manufactured in the form of m An aluminum block was manufactured through grinding, cutting, and chemical etching methods so that a total of four non-sample holes were formed in each unit area, as shown in [Figure 2].

A thermal block of the same size with no non-sample holes formed and only sample wells formed was prepared as a control. To investigate the mass reduction efficiency due to the formation of non-sample holes, the mass was measured.

As a result, as shown in [Table 1], the control 96 well thermal block with only sample wells without non-sample holes was measured at 180 g, and the 96 well thermal block of the present invention was measured at 90 g.

Therefore, it was confirmed that the heat block of the present invention reduced the mass by 50% compared to the control group. As the mass decreases, the heat capacity of the thermal block decreases, giving the advantage of being able to implement a fast ramp rate.

item Thermal Block of the present invention Control group (block without non-sampling holes) Weight of Thermal Block (96 well) 90g 180g

Example 2 Thermal uniformity measurement

Thermal uniformity was investigated by simultaneously measuring temperatures at multiple points in the thermal block and checking the deviation between those temperatures.

T-checkers that measure temperature were evenly installed in the top row, bottom row, and middle row of the heat block. The temperature was measured while increasing the temperature of the heat block from 27°C to 60°C. The temperature at each point was recorded 10 seconds after reaching 60°C. The difference between the maximum and minimum temperature values of each recorded point was calculated. The experiment was repeated more than 10 times in total to calculate the results.

As a result of the measurement, as shown in [Table 2], the thermal block of the present invention was confirmed to have a thermal uniformity of ±0.2°C. Considering that the thermal uniformity of the thermal block of other existing PCR devices is ±0.4°C, it can be seen that the thermal uniformity of the thermal block of the present invention is excellent.

item of the present invention
Thermal Block reference Thermal accuracy ±0.2℃ ±0.2℃ Thermal Block Uniformity ±0.2℃ ±0.4℃

Example 3 Ramp Rate Measurement

Reducing the mass of the block can increase heat transfer efficiency by reducing heat capacity, which can improve the speed of temperature rise or fall when the same current is supplied. To evaluate these characteristics, the temperature increase rate was compared by supplying the same current to a 16-well thermal block without a mass reduction structure and a 16-well thermal block to which the structure of the present invention was applied.

The experiment was conducted by measuring the time to reach a specific temperature. Specifically, the time to reach a given temperature was measured and compared when (1) raising the temperature from room temperature to 60°C and (2) raising the temperature from room temperature to 95°C.

As a result of the measurement, as shown in Table 3 and Figure 8, it was confirmed that the ramp rate increased in the case of the thermal block to which the mass reduction structure of the present invention was applied. Specifically, it was confirmed that the increase in ramp rate of the experimental group improved by about 16% when the temperature was raised to 95℃ and by about 34% when the temperature was raised to 60℃. These experimental results support that the temperature increase rate is improved due to the mass reduction structure of the heat block of the present invention.

Experimental conditions Ramp rate of control group (block without non-sample hole) Thermal blocks of the embodiment
ramp rate Ramp rate
improvement effect Room temperature -> 60℃ 1.3℃/sec 1.8℃/sec 34% Room temperature -> 90℃ 1.3℃/sec 1.5℃/sec 16%

100: thermal block (sample block; thermal block)
101: top surface
102: bottom surface
103: sample well
104: opening of sample well
105: floor of sample well
106: non sample hole
107: opening of non sample hole
130: unit region

Claims (8)

A thermal block for performing multiple reactions,
The thermal blocks are parallel to each other and have upper and lower surfaces having a length and width,
A plurality of sample wells open to the upper surface are formed on the upper surface, and the plurality of sample wells are arranged regularly, and
Two or more non-sample holes open to the upper surface are formed on the upper surface, and the upper surface opening area of each non-sample hole is smaller than the upper surface opening area of the sample well,
A thermal block, characterized in that two or more non-sample holes are included within a unit area defined by connecting the center points of four adjacent sample wells in the thermal block.
The heat block according to claim 1, wherein the sample well is formed to accommodate a reaction vessel.
The thermal block according to claim 1, wherein a unit area defined by connecting the center points of four adjacent sample wells in the thermal block includes four or more non-sample holes.
The method of claim 1, wherein the regular arrangement of the plurality of sample wells is a rectangular array, and a diagonal line formed by connecting the center points of four adjacent sample wells forming the unit area is at least within the unit area. A thermal block characterized by not touching or intersecting a single non-sample hole.
The heat block according to claim 1, wherein the non-sample hole is formed so that the heat conduction influence of the non-sample hole within the unit area for each of the four adjacent sample wells is the same.
The thermal block according to claim 4, wherein all straight lines connecting the center points of four adjacent sample wells forming the unit area do not contact or intersect non-sample holes within the unit area.
The thermal block according to claim 1, wherein the number of non-sample holes is greater than the number of sample wells. The thermal block according to claim 1, wherein the non-sample holes in the unit area are equal to or symmetrical with respect to the center point of each of the four sample wells forming the unit area.

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