201231646 六、發明說明： 【發明所屬之技術領域】 本發明與聚合酶連鎖反應(PCR)有關，特別是有關於一 種聚合酶連鎖反應之溫度設定方法及裝置。 【先前技術】201231646 VI. Description of the Invention: [Technical Field] The present invention relates to a polymerase chain reaction (PCR), and more particularly to a temperature setting method and apparatus for a polymerase chain reaction. [Prior Art]
聚合酶連鎖反應(PCR)用於擴增特定DNA片段，被廣 泛地應用在醫學及生物學之實驗室。此項技術一般由 20〜35個熱循環組成，每個循環包含以下三個不同溫度條 件之步驟：於95t下進行DNA變性反應(Denaturation)、 於55°C下進行引子黏合反應(Annealing)、以及於72°C下進 行延展反應(Extension)。一般聚合酶連鎖反應(pCR)係於一 熱循環設備中進行，該熱循環設備藉由調整一介質（包括氣 體、液體或固體)之溫度來改變毛細試管中整體混合液之溫 度’來進行聚合酶連鎖反應。 然而，傳統熱循環設備於各步驟間的溫度轉換皆需要 =當^ ’才歧變介質溫度錢齡賴㈣達到熱平 =導致反應過程耗時冗長；再者，由於傳統 = 显控裝置改變介質之溫度，導 【發明内容】 之溫度設 —本發明之目的在於提供一種聚合酶連鎖反應 疋方法’可縮短聚合酶連鎖反應所需之時間。 201231646 本發明之另一目的在於提供一種聚合酶連鎖反應之溫 度设疋裝置，其成本低廉，極具市場潛力。 為達上揭之目的，本發明所提供之聚合酶連鎖反應之 溫度設定方法，其包含有下列步驟：（a)將一裝有混合液 (mixture)與DNA樣品之試管置於一密閉殼體内，以及(b) 以一加熱器將位於該試管底部之混合液加熱至變性溫度 (denaturing temperature)。藉由此溫度設定方法，可縮短聚 合酶連鎖反應所需之時間。 另外，本發明所提供聚合酶連鎖反應之溫度設定裝 置，其包含有一密閉殼體，具有一用以容置該試管之容置 空間以及一位於該密閉殼體内之加熱器，該加熱器係用以 加熱位於該試管底部之混合液。該裝置結構簡單且成本低 廉’極具市場潛力。 【實施方式】 為了更瞭解本發明之特點所在’茲舉以下二較佳實施 例並配合圖式說明如下，其中： 第一圖係本發明第一較佳實施例之剖視圖； 第二圖係本發明第一較佳實施例之局部立體圖；以及 第三圖係本發明第二較佳實施例之局部立體圖。 清參閱第一圖及第二圖，係本發明第一較佳實施例所 提供聚合酶連鎖反應之溫度設定裝置10，用以設定複數試 管τ中混合液之溫度，該裝置1〇係由一密閉殼體2〇、一 加熱器30以及一絕熱層40組成。 201231646 該密閉殼體20具有一上蓋21、一下蓋23以及一由該 上蓋21與下蓋23共同界定出之容置空間25，用以供該等 試管T容置。該密閉殼體20係可隔絕外界氣流，避免外 界氣流影響該密閉殼體2〇内之溫度。 該加熱器30設於該密閉殼體20中，係由具有良好熱 傳導性之金屬製成，該加熱器30具有複數個由該加熱器 30之側面向内凹設之半圓柱狀容槽31，用以供該試管τPolymerase chain reaction (PCR) is used to amplify specific DNA fragments and is widely used in laboratories in medicine and biology. The technique generally consists of 20 to 35 thermal cycles, each cycle comprising the following three different temperature conditions: DNA denaturation at 95t, and Annealing at 55°C. And an extension reaction was carried out at 72 °C. The general polymerase chain reaction (pCR) is carried out in a thermal cycler that performs polymerization by adjusting the temperature of a medium (including gas, liquid or solid) to change the temperature of the overall mixture in the capillary tube. Enzyme chain reaction. However, the temperature conversion of the traditional thermal cycle equipment between the various steps needs to be = when the temperature of the medium is ambiguous (4) reaches the heat level = causing the reaction process to take time and tedious; moreover, because the traditional = display control device changes the medium Temperature, the temperature setting of the present invention - the object of the present invention is to provide a polymerase chain reaction reaction method "the time required to shorten the polymerase chain reaction. 201231646 Another object of the present invention is to provide a temperature setting device for a polymerase chain reaction which is low in cost and has great market potential. For the purpose of the above disclosure, the temperature setting method of the polymerase chain reaction provided by the present invention comprises the following steps: (a) placing a test tube containing a mixture and a DNA sample in a closed casing. Inside, and (b) heating the mixture at the bottom of the tube to a denaturing temperature with a heater. By this temperature setting method, the time required for the polymerase chain reaction can be shortened. In addition, the temperature setting device for the polymerase chain reaction provided by the present invention comprises a sealed casing having a receiving space for accommodating the test tube and a heater located in the sealed casing. Used to heat the mixture at the bottom of the tube. The device is simple in structure and low in cost. It has great market potential. BRIEF DESCRIPTION OF THE DRAWINGS In order to better understand the features of the present invention, the following two preferred embodiments are described below with reference to the accompanying drawings, wherein: FIG. 1 is a cross-sectional view of a first preferred embodiment of the present invention; A partial perspective view of a first preferred embodiment of the invention; and a third perspective view of a second preferred embodiment of the invention. Referring to the first and second figures, a temperature setting device 10 for a polymerase chain reaction according to a first preferred embodiment of the present invention is used to set the temperature of a mixed solution in a plurality of test tubes τ. The sealed casing 2, a heater 30, and a heat insulating layer 40 are composed. 201231646 The sealed casing 20 has an upper cover 21, a lower cover 23 and an accommodation space 25 defined by the upper cover 21 and the lower cover 23 for receiving the test tubes T. The sealed casing 20 is insulated from the outside air flow to prevent the external airflow from affecting the temperature inside the sealed casing 2. The heater 30 is disposed in the sealed casing 20 and is made of a metal having good thermal conductivity. The heater 30 has a plurality of semi-cylindrical cavities 31 recessed inwardly from the side of the heater 30. Used for the test tube τ
底部容置，且各該容槽31具有一假想軸線32垂直或約垂 直於一水平面。該加熱器30可依需要設定其溫度於約 90〜98°C，並透過熱傳導加熱位於該試管τ底部之混合液 至變性溫度(denaturing temperature) 〇 該絕熱層40係覆蓋於該加熱器30之頂面，該絕熱層 4〇具有複數個鏤空部4卜其位置分別對應於該加熱器3〇 之該等容槽3卜用以供該試管τ穿入。該絕熱層4〇可阻 絕該加熱器30之熱能，使位於該絕熱層4〇上方之試管τ 内的混合液溫度不致受該加熱器3〇影響。 實際操作該裝置時，可將複數個財混合液⑽餘) 與DNA樣品之試管T置放於―固找c(此為習知技術， 容不贅述)上，使料試管T穿過魏_⑽之鏤空部41 而^底端容置於該加熱器3〇的容槽31 +，該加熱器％ 可將…式管T底部之混合液加熱至變性溫度， 90〜贼’於本實施例中為約95ΐ，如此，該試管T中渴 峨而產生熱對流’致使位於底部之混合浴 向上流動歧漸降溫。料言之’1如鱗，試管「 201231646 液面溫度約等同於室溫(約叫隨著試管τ 二=的液面溫度也會逐漸上升，此時，試管： 即可進行引的黏合溫度(如抑)，於此 逼付子之黏合，並且，試管了中的混合液 一^因溫度梯度錢㈣子進聽展反料溫度（約 )，於此即可進概狀應，而纽合液_向下對流 至試管Τ底科，溫度到達95t，即可再錢行變性反應， 如此不斷《，達祕軸健狀DNA細以供檢測 用之目的。當然、，隨著不斷的熱循環，混合液的液面溫度 會逐漸升S至55。(：以上，超過引子轉合溫度，但事實 上’早在溫度升高至坑以前，混合賴賴環次數已足 夠使聚合酶連鎖反應絲。藉此，可大幅雜傳統方法變 換溫度所需之時間，且應用此方法之溫度設定裝置結構簡 單，製造成本低廉，極具市場潛力。 於本發明中，前述之混合液包含有緩衝溶液(buffer)、 驗基對(dNTPs)、聚合酶(Taqp〇iymerase)及引子(讲如⑽）。 依據本發明之精神，前述實施例中加熱器之結構可加 以變化，如第三圖所示，為本發明第二較佳實施例所提供 之溫度設定裝置，其中’加熱器50之容槽51係由該加熱 器50之頂面向下凹設而呈圓柱狀。事實上，該加熱器之加 熱方式除了前揭之熱傳導方式之外，亦可依需要而變化為 以熱輻射的方式對混合液進行加熱，例如透過微波方式予 以加熱。 201231646 在此，需補充說明的是，該加熱器之容槽的數目，可 依需要設一個或一個以上，與其對應之絕熱層之鏤空部的 數目亦同。 本發明於前揭實施例中所揭露之構成元件，僅為舉例 說明，並非用來限制本案之範圍，其他等效元件的替代或 變化，亦應為本案之申請專利範圍所涵蓋。 201231646 【圖式簡單說明】 第一圖係本發明第一較佳實施例之剖視圖； 第二圖係本發明第一較佳實施例之局部立體圖；以及 第三圖係本發明第二較佳實施例之局部立體圖。 21上蓋 25容置空間 31容槽 40絕熱層 50加熱器 T試管The bottom portion is received, and each of the pockets 31 has an imaginary axis 32 that is perpendicular or about perpendicular to a horizontal plane. The heater 30 can be set to a temperature of about 90 to 98 ° C as needed, and heat the mixture at the bottom of the tube τ to a denaturation temperature by heat conduction. The heat insulating layer 40 covers the heater 30. In the top surface, the heat insulating layer 4 has a plurality of hollow portions 4, the positions of which correspond to the heat sinks 3 of the heaters 3, respectively, for the test tube τ to penetrate. The heat insulating layer 4 阻 can block the heat energy of the heater 30 so that the temperature of the mixed liquid in the test tube τ located above the heat insulating layer 4 is not affected by the heater 3 . When the device is actually operated, a plurality of the mixed liquids (10) and the test tube T of the DNA sample may be placed on the "solid-seeking c" (this is a conventional technique, which is not described above), so that the test tube T passes through the Wei_ (10) the hollow portion 41 and the bottom end accommodates the tank 31 + of the heater 3, the heater % can heat the mixture of the bottom of the tube T to the denaturation temperature, 90~ thief' in this embodiment The middle is about 95 ΐ, so that the tube T is thirsty and produces heat convection, causing the mixing bath at the bottom to gradually cool down. It is said that '1 scale, test tube '201231646 liquid level temperature is about equal to room temperature (about the same as the test tube τ 2 = the liquid surface temperature will also gradually rise, at this time, the test tube: can be used to conduct the bonding temperature ( If it is suppressed, the binder is bonded to this, and the mixture in the test tube is due to the temperature gradient (4), and the temperature is (about). The liquid _ convection downwards to the test tube, the temperature reaches 95t, then the denaturation reaction can be carried out again, so that the DNA is fine for the purpose of detection. Of course, with the continuous thermal cycle The liquid level of the mixture will gradually increase by S to 55. (: Above, exceed the primer transfer temperature, but in fact 'before the temperature rises to the pit, the number of mixed lysing rings is enough to cause the polymerase chain reaction Therefore, the time required for changing the temperature by the conventional method can be greatly increased, and the temperature setting device using the method has the advantages of simple structure, low manufacturing cost, and great market potential. In the present invention, the foregoing mixed solution contains a buffer solution ( Buffer), base pair (dNTPs), polymerase ( Taqp〇iymerase) and primer (refer to (10)). According to the spirit of the present invention, the structure of the heater in the foregoing embodiment may be changed, as shown in the third figure, the temperature provided by the second preferred embodiment of the present invention. The setting device, wherein the receptacle 51 of the heater 50 is recessed from the top surface of the heater 50 to have a cylindrical shape. In fact, the heating method of the heater may be in addition to the heat conduction method disclosed above. If necessary, the mixture is heated by means of heat radiation, for example, by microwave heating. 201231646 Here, it should be additionally noted that the number of the tanks of the heater may be one or more as needed. The number of the hollow portions of the heat insulating layer is the same as that of the corresponding heat insulating layer. The constituent elements disclosed in the foregoing embodiments are merely illustrative and are not intended to limit the scope of the present invention. The present invention is covered by the scope of the patent application. 201231646 [Simplified illustration of the drawings] The first drawing is a cross-sectional view of the first preferred embodiment of the present invention; A partial perspective view of the embodiment; FIG line and a third partial perspective view of a second preferred embodiment of the present invention, the upper cover 25 accommodating space 21 accommodating groove 40 insulating layer 31 50 T heater tubes.
【主要元件符號說明】 ίο溫度設定裝置 20密閉殼體 23下蓋 30加熱器 32假想軸線 41鏤空部 51容槽 C固定架[Description of main component symbols] ίο Temperature setting device 20 Hermetic housing 23 Lower cover 30 Heater 32 Imaginary axis 41 hollow part 51 Capacity C holder