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Micro-fluidic PCR chip

A microfluidic and chip technology, which is applied in the field of microfluidics, can solve the problems of difficult processing, plastic PCR chips are easy to burst, and burst, so as to reduce the risk of easy burst, avoid chip easy burst, and increase the effect of thermal conductivity

Pending Publication Date: 2021-04-20
BIOISLAND LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above patents all provide microfluidic PCR chips, since the temperature control element is only on one side of the chip, the temperature changes slowly, and it is easy to cause uneven heating of the liquid in the flow channel.
In addition, the temperature distribution of the chip is not symmetrical, which can easily lead to deformation or bursting of the chip, especially when the chip material is plastic
At the same time, setting the reaction flow channel on the same side of the chip will also result in a narrow and long chip size, which is difficult to process and increases the difficulty of preparation.
[0005] Therefore, it is hoped to provide a new microfluidic PCR chip, which can reduce the thickness of the chip, reduce the difficulty of production, and solve the shortcomings of the unstable and inaccurate temperature of the PCR chip, and the plastic PCR chip is easy to burst.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] This embodiment provides a microfluidic PCR chip.

[0051] Such as figure 1 As shown, the microfluidic PCR chip includes a set of thermal cycle reaction parts 1 and a plurality of temperature control devices 2 and drive devices located on both sides of the thermal cycle reaction part 1 .

[0052] Such as figure 2 As shown, the heat cycle reaction part 1 is provided with a heat cycle flow channel 11, and the heat cycle flow channel 11 is arranged in a serpentine shape, including 36 single cycle flow channels connected from end to end and symmetrically distributed. The road has a high-temperature section and a low-temperature section. A plurality of high-temperature sections form a high-temperature section 112, and a plurality of low-temperature sections form a low-temperature section 111. The low-temperature section 111 is symmetrically distributed on both sides of the high-temperature section 112. The length of the high-temperature section is The length ratio of the ...

Embodiment 2

[0056] This embodiment provides a method for performing a PCR reaction using the microfluidic PCR chip provided in Embodiment 1.

[0057] The sample enters the preheating flow channel from the sample injection flow channel, and the sample is preheated by setting the temperature of the preheating flow channel at 95°C. After the preheating is completed, it enters the circulation flow channel connected to the preheating flow channel for thermal cycle. Set the low temperature to 60°C, so that the sample can go through the circulation flow of high temperature (95°C)-low temperature (60°C)...high temperature-low temperature. During the thermal cycle, the sample can pass through the observation window formed between the MCH ceramic heating elements Observe the reaction situation, and after the cycle is completed, the sample is discharged through the sample discharge channel.

Embodiment 3

[0059] This embodiment provides a method for performing a PCR reaction using the microfluidic PCR chip provided in Embodiment 1.

[0060] Use 2×PCR Mix, double-stranded DNA dye (20×), template pGFP-C-shLenti, the sequence is shown in SEQ ID No:1;

[0061] Primer TGFP-128-s: TTCACCGACAAGATCATCC;

[0062] TGFP-128-anti:ACCACGGAGCTGTAGTAG.

[0063] The reaction solution preparation method is shown in Table 1:

[0064] Table 1

[0065] Element Content / μL 2×PCR Mix 15 TGFP-128-s (10μM) 1.5 TGFP-128-anti (10μM) 1.5 pGFP-C-shLenti (200ng / μL) 0.5 Double-stranded DNA dye (20×) 7.5 water 24

[0066] Sample flow rate: 0.5μL / min, hot start temperature 95°C, time 10min, high temperature time at 95°C for each cycle: 10s, low temperature time at 60°C for each cycle: 15s.

[0067] image 3 is the fluorescence map of the sample after preheating and before thermal cycling, Figure 4 It is the fluorescence diagram of the sample after 35 t...

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Abstract

The invention provides a micro-fluidic PCR chip. The micro-fluidic PCR chip comprises at least one group of thermal cycle reaction parts, wherein the thermal cycle reaction parts are provided with thermal cycle flow channels, the thermal cycle flow channels are arranged in a snake shape and comprise 2n single cycle flow channels which communicate with end to end and are symmetrically distributed, and each single cycle flow channel comprises a high-temperature section and a low-temperature section, so that a sample is subjected to high-temperature and low-temperature alternate cycle reaction in the single cycle flow channels. According to the micro-fluidic PCR chip, the high-temperature and low-temperature alternate cycle reaction of the sample in the thermal cycle flow channels can be realized, and the chip can reduce the thicknesses of the thermal cycle flow channels, increase the strengths of the thermal cycle flow channels, improve the heat-conducting property of the thermal cycle flow channels and improve the thermal conductivity of the thermal cycle flow channels. The risk of bursting of the heat cycle flow channel due to non-uniform heating of the two sides can be avoided, and the chip manufacturing difficulty can be reduced through the structure.

Description

technical field [0001] The invention belongs to the field of microfluidic technology, and relates to a microfluidic PCR chip. Background technique [0002] PCR is currently the most common nucleic acid amplification reaction, which can specifically amplify specific low-concentration double-stranded deoxyribonucleotide (Deoxyribonucleic acid, DNA) fragments within one or two hours in vitro. PCR is a technology that can be used in various occasions such as genetic analysis, medical diagnosis, food safety, and forensic identification. [0003] The classic PCR technique mainly includes 5 steps: pretreatment (sample treatment, nucleic acid extraction and reagent addition, etc.), high temperature denaturation (DNA denaturation to form two single strands, ~95°C), low temperature annealing (DNA single strand and primer annealing, 55-70°C), temperature-optimized extension (substrand extension DNA doubling, ~72°C) and post-processing (collection and electrophoresis detection, etc.). ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/38C12M1/02C12M1/00
Inventor 董鸣林至诚
Owner BIOISLAND LAB