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WO2016117958A1 - Dispositif de réglage de la température pour un équipement d'analyse d'adn - Google Patents

Dispositif de réglage de la température pour un équipement d'analyse d'adn Download PDF

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Publication number
WO2016117958A1
WO2016117958A1 PCT/KR2016/000691 KR2016000691W WO2016117958A1 WO 2016117958 A1 WO2016117958 A1 WO 2016117958A1 KR 2016000691 W KR2016000691 W KR 2016000691W WO 2016117958 A1 WO2016117958 A1 WO 2016117958A1
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WO
WIPO (PCT)
Prior art keywords
mounting holes
temperature
heaters
dna sample
turned
Prior art date
Application number
PCT/KR2016/000691
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English (en)
Korean (ko)
Inventor
이진우
송재영
신연경
김은주
조수동
박최규
김희정
김은미
Original Assignee
(주)미코바이오메드
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Application filed by (주)미코바이오메드 filed Critical (주)미코바이오메드
Publication of WO2016117958A1 publication Critical patent/WO2016117958A1/fr

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples

Definitions

  • the present invention relates to a temperature control device for DNA analysis equipment, and more particularly to a device for heating the DNA sample to a certain temperature in order to amplify the DNA sample to enable analysis.
  • PCR polymerase chain reaction
  • the PCR method generally comprises the steps of decomposing the double helix structure of the collected DNA (denaturation, 90 ⁇ 96 °C), conjugating the primer (annealing, 40 ⁇ 65 °C), replication to the original double helix structure It comprises the step (polymerization, 70 ⁇ 74 °C). In these steps, it is important to uniformly and precisely control each temperature for heating or cooling the collected DNA.
  • the PCR method includes a lower heater including a heater for heating the tubes around the mounting holes while having a plurality of mounting holes in which the collected DNA is loaded on the upper and the upper portion of the lower heater.
  • a heating apparatus including an upper heater coupled with the lower heater is used to cover the mounting holes.
  • the heating device has a structure in which the heater heats the whole surrounding the mounting holes. Therefore, the heat transfer distance from the heater is farther from the center area of the mounting holes than the outer area of the mounting holes. Therefore, there is a problem in that a relatively large amount of time is consumed in order to uniformly and precisely adjust the temperature of the DNA contained in the tubes according to the positions of the tubes at the temperature according to each step of the PCR method.
  • the present invention provides a temperature control device capable of controlling the temperature of the DNA sample uniformly and precisely and very quickly in amplifying the DNA sample for analysis.
  • the temperature control device for DNA analysis equipment comprises a lower heater and an upper heater.
  • the lower heater has a plurality of mounting holes in which a tube containing a DNA sample for analysis is mounted on the upper side of the lower heater, arranged side by side along a plurality of rows, and located at both sides of each of the mounting holes corresponding to each column. Heaters.
  • the upper heater is coupled to the lower heater to cover the mounting holes on the upper portion of the lower heater.
  • the distance between the rows of the mounting holes is equal to the distance between the mounting holes in the same row or longer than the distance between the mounting holes in the same row depending on the size of the first and second heaters. Can be.
  • the lower heater may further include a cooling cooler positioned below the mounting holes for additional temperature control.
  • the cooling cooler may include a peltier device.
  • the control unit may further include.
  • the heating temperature may be set in advance.
  • the temperature control unit When one of the first and second heaters is turned on by the temperature control unit and the cooling cooler is turned on, the DNA sample and primer contained in the tube are combined. It may be characterized in that the cooling temperature is preset.
  • the upper heater may have opening holes corresponding to each of the mounting holes to be exposed upward.
  • the temperature control device is for checking the optical state of the fluorescent material, for example, the luminescence state, which is included when the DNA sample contained in the tube mounted in each of the mounting holes is replicated through the opening holes on the upper heater.
  • the optical module may further include.
  • the temperature control device for DNA analysis equipment of the present invention in a state in which a plurality of mounting holes in which a tube containing a DNA sample for analysis is mounted are arranged side by side along a plurality of rows, the amount of mounting holes corresponding to each column.
  • the heat transfer distance between the DNA sample contained in the tube and the first and second heaters and the cooling cooler is constant. It can be shortened very close.
  • the DNA sample can be uniformly and precisely adjusted according to the position at the temperature according to each step in the amplification process described in the background art, and the time for controlling the temperatures due to the shortening of the heat transfer distance is also remarkable. Can be shortened.
  • FIG. 1 is a perspective view schematically showing a temperature control device according to an embodiment of the present invention.
  • FIG. 2 is a view showing in detail the lower heater of the temperature control device shown in FIG.
  • FIG. 3 is a side view of the mounting holes shown in FIG. 2.
  • 4 to 7 are graphs experimentally showing the temperature distribution of the mounting holes controlled by the lower heater shown in FIGS. 2 and 3.
  • FIG. 8 is a view showing in detail the upper heater of the temperature control device shown in FIG.
  • a plurality of mounting holes in which a tube containing a DNA sample for analysis is mounted is arranged side by side along a plurality of rows, and mounting holes corresponding to each column.
  • a lower heater including first and second heaters positioned at both sides of the field, and an upper heater coupled to the lower heater to cover the mounting holes on the upper portion of the lower heater.
  • first and second may be used to describe various components, but the components should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from another.
  • the first component may be referred to as the second component, and similarly, the second component may also be referred to as the first component.
  • FIG. 1 is a perspective view schematically showing a temperature control device according to an embodiment of the present invention
  • Figure 2 is a view showing a lower heater of the temperature control device shown in Figure 1 in detail
  • Figure 3 is shown in Figure 2 Side view of the mounting holes.
  • the temperature control device 100 heats or cools the DNA sample in an analysis device that amplifies and analyzes a DNA sample according to the PCR method described in the background art. Used to adjust to a specific temperature.
  • the temperature control device 100 includes a lower heater 200 and an upper heater 300.
  • the lower heater 200 has a plurality of mounting holes 210 thereon.
  • the mounting holes 210 are arranged side by side along a plurality of rows.
  • the mounting holes 210 may be arranged side by side along three first, second and third rows R1, R2, and R3.
  • Each of the mounting holes 210 is equipped with a tube 10 containing the DNA sample.
  • the guide groove 220 is formed on the lower heater 200, and the PCR plate 20 may be partially inserted into the guide groove 220 so that the PCR plate 20 is stably placed. have.
  • at least one corner of the PCR plate 20 and the guide groove 220 may be formed with first and second alignment units 22 and 222 for aligning the positions.
  • the first and second alignment units 22 and 222 may have a rounded structure as shown in FIGS. 1 and 2.
  • kits connected to the tubes 10 may be used to correspond to any one of the mounting holes 210.
  • the kit may have a structure in which eight tubes 10 commonly used in the art for analyzing the DNA sample are connected in a line.
  • the lower heater 200 includes first and second heaters 230 and 240 bonded to heat DNA samples contained in each of the tubes 10 on both sides of the mounting holes 210 corresponding to each row. Include.
  • the plurality of first and second heaters 230 and 240 may be disposed to be long along each row of the mounting holes 210.
  • the distance G1 between the columns of the mounting holes 210 indicates that the DNA samples contained in each of the tubes 10 are thermally interfered by adjacent first or second heaters 230 and 240. In order to prevent it, it must be secured longer than or equal to the distance G2 between the mounting holes 210 according to the same row.
  • the lower heater 200 may further include a cooling cooler 250 bonded to cool the DNA samples contained in each of the tubes 10 under the mounting holes 210.
  • the cooling cooler 250 may include a peltier device capable of precisely adjusting the cooling temperature of the DNA samples contained in each of the tubes 10 according to a potential difference using semiconductor characteristics.
  • the lower heater 200 including the first and second heaters 230 and 240 and the cooling cooler 250 may step the DNA sample in each step when amplifying the DNA sample according to the PCR method described in the background. Each temperature is precisely adjusted to different temperatures. Accordingly, the temperature control device 100 may include a temperature controller 400 connected to the first and second heaters 230 and 240 and the cooling cooler 250 to precisely control them. .
  • the temperature controller 400 turns on / off the first and second heaters 230 and 240 and the cooling cooler 250 at preset heating and cooling temperatures, respectively. ) Is adjusted to a different temperature for each step according to the PCR method.
  • the temperature control unit 400 will be described in detail with respect to a method of adjusting to a temperature set differently for each step in the PCR method.
  • the heating temperature is preset to decompose the structure, and when any one of the first and second heaters 230 and 240 is turned on while the cooler 250 is off.
  • the heating temperature is preset such that a primer is coupled to a DNA sample contained in each of the tubes 10.
  • a DNA sample contained in each of the tubes 10 is duplicated.
  • the heating temperature is preset to replicate in a spiral structure.
  • the temperature control unit 400 decomposes the double-helical structure of the DNA sample in the PCR method, in which the cooling cooler 250 is turned off, and the first and second heaters 230 and 240.
  • the first and second heaters 230 and 240 in the step of heating the DNA sample and adding a primer to the DNA sample to bind to the decomposed double helix structure.
  • the DNA sample is duplicated in a double helix structure in the step of turning off the cooling cooler 250 (off) ), The DNA sample may be heated again by turning on any one of the first and second heaters 230 and 240.
  • the temperature controller 400 By simply controlling the first and second heaters 230 and 240 and the cooling cooler 250 on / off, thereby controlling the temperature of the DNA sample to the predetermined heating and cooling temperatures. I can regulate it. Therefore, the temperature of the DNA sample can be controlled simply and quickly to a temperature set differently for each step according to the PCR method, and can significantly shorten the amplification time of the DNA sample according to the PCR method.
  • the method described in the background art is used. While it takes about 1 hour, using the temperature control unit 400 of the present invention can achieve an effect of shortening the time by about 60% or more in about 20 minutes.
  • 4 to 7 are graphs experimentally showing the temperature distribution of the mounting holes controlled by the lower heater shown in FIGS. 2 and 3.
  • DNA double helix degradation temperature, primer binding temperature, DNA double helix replication temperature were set to 94 °C, 55 °C, 72 °C, respectively.
  • the cooling cooler 250 is turned off to heat the DNA sample contained in each of the tubes 10 to about 94 ° C. through the temperature controller 400.
  • a temperature range of each position along a line of each of P1 to P4 of the mounting holes 210 is less than or equal to 0.03 ° C. or less. It could be confirmed that appears.
  • the temperature range according to the position of the mounting holes 210 is only 0.2 ° C. or lower, a considerable precision is maintained. It can be demonstrated that the uniformity and precision are at a considerable level.
  • the amount of the mounting holes 210 corresponding to the respective columns Bonding the cooling cooler 250 to the lower portion of the mounting holes 210 while joining the first and second heaters 230 and 240 to each of the sides, so that the DNA sample and the first and second heaters 230, The heat transfer distance between the 240 and the cooling cooler 250 may be shortened to a very close and constant distance.
  • a temperature according to each step in the amplification process described in the background for example, 94 °C, 55 °C and 72 °C.
  • FIG. 8 is a view showing in detail the upper heater of the temperature control device shown in FIG.
  • the upper heater 300 is coupled to the lower heater 200 to cover the mounting holes 210 on the upper portion of the lower heater 200.
  • the upper heater 300 may be coupled to the lower heater 200 while having a plate shape to be mounted on the mounting holes 210 to cover the tubes 10 containing the DNA sample.
  • a cover film (not shown) may be attached to the open upper portions of the tubes 10 to preferentially prevent evaporation of moisture from the DNA sample when mounted in each of the mounting holes 210.
  • the upper heater 300 always heats the upper portion of the DNA sample at about 90 to 100 ° C. during the steps according to the PCR method, thereby preventing moisture from evaporating from the DNA samples contained in each of the tubes 10. It can prevent.
  • the upper heater 300 may further include a second guide groove 310 on the lower surface. The second guide groove 310 may guide the position of the PCR plate 20 when the lower heater 200 is covered.
  • the temperature control device 100 detects in real time the optical state of the fluorescent material contained in the DNA samples contained in each of the tubes 10, in particular the emission state during the amplification process according to the PCR method
  • the optical module 500 may further include.
  • the fluorescent material is added when the primer is added during the PCR method.
  • the optical module 500 is formed in the state in which the opening holes 320 corresponding to each of the mounting holes 210 are formed in the upper heater 300 to expose the mounting holes 210 upward.
  • the DNA sample is scanned every time the PCR method is performed through the opening holes 320 in the upper portion of the upper heater 300 to detect the emission intensity of the fluorescent material included in the DNA sample. Therefore, it is possible to confirm in real time whether the DNA sample can be analyzed even in the course of the PCR method.
  • both sides of the mounting holes corresponding to the columns are arranged in parallel with the plurality of mounting holes in which the tubes containing the DNA sample for analysis are mounted.
  • the first and second heaters are bonded to each other, and a cooling cooler is attached to the lower portion of the mounting holes. Therefore, the temperature control device can be utilized to dramatically shorten the amplification time of the DNA sample by controlling the DNA sample uniformly, precisely and quickly at different temperatures when amplifying the DNA sample according to the PCR method.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne un dispositif de réglage de la température pour un équipement d'analyse d'ADN comprenant un dispositif de chauffage inférieur et un dispositif de chauffage supérieur. Le dispositif de chauffage inférieur comprend : une pluralité de trous de montage qui sont disposés côte à côte le long d'une pluralité de colonnes sur la partie supérieure du dispositif de chauffage inférieur, la pluralité des trous de montage présentant des tubes montés dessus qui maintiennent les échantillons d'ADN à analyser; des premier et second dispositifs de chauffage qui sont positionnés sur chacun des deux côtés des trous de montage correspondant à chaque colonne; et un dispositif de refroidissement sur la partie inférieure. Le dispositif de chauffage supérieur s'accouple au dispositif de chauffage inférieur afin que les trous de montage sur la partie supérieure du dispositif de chauffage inférieur soient recouverts.
PCT/KR2016/000691 2015-01-22 2016-01-22 Dispositif de réglage de la température pour un équipement d'analyse d'adn WO2016117958A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020150010380A KR101698996B1 (ko) 2015-01-22 2015-01-22 Dna 분석 장비용 온도 조절 장치
KR10-2015-0010380 2015-01-22

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WO2016117958A1 true WO2016117958A1 (fr) 2016-07-28

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Publication number Priority date Publication date Assignee Title
KR101882239B1 (ko) * 2016-12-06 2018-07-26 (주)옵토레인 다중 온도설정이 가능한 피씨알모듈, 이를 포함하는 피씨알시스템, 및 이를 이용한 피씨알실험방법
KR102126520B1 (ko) * 2020-02-12 2020-06-24 주식회사 케이에이치메디칼 핵산 증폭 장치
KR102434822B1 (ko) * 2021-10-06 2022-08-22 주식회사 위즈바이오솔루션 분자 진단용 고속 열순환이 가능한 pcr 장치

Citations (5)

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Publication number Priority date Publication date Assignee Title
JP2002536155A (ja) * 1999-02-05 2002-10-29 ビラテック ゲゼルシャフト ツア エントヴィックルング ビオテヒノロギッシヤー ジステーメ ミット ベシュレンクテル ハフツング 個々の容器を選択的に温度調整するための装置
KR100803090B1 (ko) * 2006-02-27 2008-02-18 주식회사 엘지화학 촉매 탐색용 반응 장치
WO2011021640A1 (fr) * 2009-08-20 2011-02-24 タカラバイオ株式会社 Dispositif de cyclage thermique
JP5019300B2 (ja) * 2006-04-13 2012-09-05 大学共同利用機関法人情報・システム研究機構 マルチウェルインキュベーション装置及びこれを用いた分析方法
JP5619690B2 (ja) * 2011-07-26 2014-11-05 株式会社古河電工アドバンストエンジニアリング 蛍光測定装置、ウェルの製造方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101040489B1 (ko) 2008-07-16 2011-06-09 연세대학교 산학협력단 실시간 모니터링이 가능한 ｐｃｒ 장치

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002536155A (ja) * 1999-02-05 2002-10-29 ビラテック ゲゼルシャフト ツア エントヴィックルング ビオテヒノロギッシヤー ジステーメ ミット ベシュレンクテル ハフツング 個々の容器を選択的に温度調整するための装置
KR100803090B1 (ko) * 2006-02-27 2008-02-18 주식회사 엘지화학 촉매 탐색용 반응 장치
JP5019300B2 (ja) * 2006-04-13 2012-09-05 大学共同利用機関法人情報・システム研究機構 マルチウェルインキュベーション装置及びこれを用いた分析方法
WO2011021640A1 (fr) * 2009-08-20 2011-02-24 タカラバイオ株式会社 Dispositif de cyclage thermique
JP5619690B2 (ja) * 2011-07-26 2014-11-05 株式会社古河電工アドバンストエンジニアリング 蛍光測定装置、ウェルの製造方法

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KR20160090926A (ko) 2016-08-02

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