Thermal medium flowing type PCR amplification method

A heat medium and medium technology, which is applied in the field of PCR amplification to achieve the effect of avoiding perishability and strong versatility

Inactive Publication Date: 2018-07-20
冷彦宁
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In short, the above contradictions of commonly used PCR technology can only be chosen and cannot be resolved

Method used

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  • Thermal medium flowing type PCR amplification method
  • Thermal medium flowing type PCR amplification method

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Experimental program
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Effect test

Embodiment 1

[0077] Such as figure 1 As shown, the present embodiment provides a thermal medium flow PCR method for thermal medium recycling:

[0078] (1) Add the PCR reaction solution into the 0.2ml reaction container 21, and insert the reaction container into the heat medium container 22.

[0079] (2) Heat the heat medium 23 to 95° C., 50° C., and 70° C. in the three spaces of the heat medium control device 24 respectively.

[0080] (3) The heat medium control device 24 makes the heat medium 23 at 95°C flow through the heat medium container 22, and is recovered to the corresponding space by the heat medium driving device 25, and heated to 95°C. Since the reaction vessel 21 is soaked in the heat medium solution, it will soon reach 95° C. due to the heat exchange, and the nucleic acid denaturation process is completed.

[0081] (4) The heat medium control device 24 makes the heat medium 23 at 50°C flow through the heat medium container 22, and is recovered to the corresponding space by the...

Embodiment 2

[0090] This embodiment provides a kind of fast PCR equipment that can be integrated, and its implementation mode is as follows:

[0091] (1) One or more reaction vessels are integrated with the heat medium vessel. The PCR reaction liquid forms a 1mm thick liquid layer in the reaction container, and the reaction container is located in a heat medium container with an inner space of 5 mm in thickness, 10 mm in height and 70 mm in length, and the heat medium container is opened at both ends to become a heat medium passage.

[0092] (2) Heat the heat medium (air) to 95°C and 60°C respectively in the three spaces of the heat medium control device.

[0093] (3) Heat medium (air) at 95°C and 60°C enters from one end of the heat medium container and discharges from the other end as required.

[0094] (4), repeat step (3) 40 times to complete the PCR process.

[0095] The advantages of this embodiment over semiconductor heating type PCR:

[0096] (1), fast. On the one hand, the rea...

Embodiment 3

[0100] This embodiment provides a heat medium flow heat exchange system. Such as figure 2 As shown, the heat medium flow heat exchange system 1 of this embodiment includes: a temperature control device 2 , a reaction vessel 3 , a heat medium container 4 , a heat medium driver (not shown in the figure), and a heat medium channel 5 .

[0101] Wherein, the temperature control device 2 is used to obtain a plurality of fluid heat media with different temperatures. For example, for PCR reaction, three different temperatures of fluid heat medium can be provided, such as 95°C, 50°C and 70°C. It can also provide fluid heat medium with two different temperatures, such as 95°C and 65°C. Of course, the number of fluid heat mediums is not limited to two or three, and may be more, which needs to be set according to the specific type of reaction. The temperature provided is not limited to the above-mentioned temperature, and still depends on the specific type of reaction and the type of ...

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Abstract

The invention provides a thermal medium flowing type PCR amplification method. The method has the advantages that the method is efficient, quick and easy for minimization and PCR amplification can becarried out on the interior of a complex container. The method comprises the following steps: inserting or connecting a reaction container filled with a PCR reaction solution into a thermal medium container so that the reaction container can be in contact with or be soaked into a thermal medium, enabling thermal media with different temperatures to flow through the thermal medium container by a thermal medium driving device to cause that the temperature of the reaction container in contact with the thermal media changes, so that PCR amplification is completed inside the reaction container. Themethod is extremely high in reaction speed, conventional PCR operation for 1.5 h can be competed within 10 min, and PCR amplification can be carried out inside the complex container. In short, the method is high in speed, high in universality, easy for minimization and wide in range of application, can be used for temperature control of the interior of the complex container, and can be particularly applied to the fields needing high automation and minimization such as POCT.

Description

technical field [0001] The present invention mainly provides an efficient, fast and easy miniaturized PCR amplification method. It can be used in related fields such as life science research, inspection and quarantine, justice, medical diagnosis and research. Background technique [0002] Polymerase chain reaction (PCR) is the most important method of nucleic acid research, widely used in life science research, inspection and quarantine, justice, medical diagnosis and research and other related fields. [0003] PCR amplification uses different temperatures to complete the process of denaturation, renaturation, amplification and extension of nucleic acid. Repeating the above process can amplify a large number of required target gene sequences. Generally, a three-step method is used for amplification, that is, the temperature is raised to 95°C to denature and dissociate the nucleic acid in the reaction solution into single strands, the temperature is lowered to 50°C for renat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/38C12M1/00C12Q1/686
CPCB01L7/525B01L2300/1838B01L2300/185C12Q1/686C12Q2527/101
Inventor 冷彦宁
Owner 冷彦宁
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