A functionalized microfluidic chip and its method for PCR product analysis

A microfluidic chip and functional technology, applied in the field of biological analysis, can solve the problems of cumbersome operation and achieve the effect of increasing the injection volume, reducing the number of electrodes, and reducing the reaction time of PCR

Active Publication Date: 2011-12-21
GUANGZHOU IMPROVE MEDICAL TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Isotachophoresis can improve detection sensitivity through sample preconcentration, but in most cases, two buffers, leading and trailing, are required, and the operation is cumbersome

Method used

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  • A functionalized microfluidic chip and its method for PCR product analysis
  • A functionalized microfluidic chip and its method for PCR product analysis
  • A functionalized microfluidic chip and its method for PCR product analysis

Examples

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Comparison scheme
Effect test

Embodiment 1

[0039] A functionalized microfluidic chip made of PMMA, the configuration is as follows: image 3 As shown, the chip is composed of 4 reservoirs, a separation channel and a sample injection channel. The 4 reservoirs are buffer solution pool 1, buffer waste solution pool 2, sample pool 3 and sample injection waste solution pool 4. The sample pool 3 and the sample injection waste pool 4 are located on opposite sides of the separation channel. The overlapping area between the separation channel and the injection channel is the effective injection channel. The size of all channels is 50 μm in width and 20 μm in depth, the effective sampling channel is 1.9 cm long, and the inner surface of the chip channel is not modified. The trailing dielectric medium is N-(ethyl-hydroxyethyl)piperazine-N-2ethanesulfonic acid (HEPES), and the DNA sieving medium is 3% (mass volume percentage, the unit is g / 100ml) hydroxyethyl cellulose. The sample is Φx174 HaeIII Digest, the total DNA content is...

Embodiment 2

[0046] The functionalized microfluidic chip configuration of the glass material used is as follows: Figure 5 As shown, the chip is composed of 4 reservoirs, a separation channel and a sample injection channel. The 4 reservoirs are buffer solution pool 1, buffer waste solution pool 2, sample pool 3 and sample injection waste solution pool 4. The sample pool 3 and the sample injection waste pool 4 are located on the same side of the separation channel. The overlapping area between the separation channel and the injection channel is the effective injection channel. The size of the chip channel is 50 μm in width and 20 μm in depth, the length of the effective sampling channel is 1.9 cm, and the width of the effective sampling channel is 50 μm. The inner surface of the chip channel is modified with linear polyacrylamide. Use this chip to analyze a PCR product of 120 base pairs, and use the DL2000 DNA standard sample as an internal reference. The buffer contained 20mmol / L MOPS a...

Embodiment 3

[0053] A functionalized microfluidic chip made of PDMS material, the configuration is as follows: image 3 As shown, the chip is composed of 4 reservoirs, a separation channel and a sample injection channel. The 4 reservoirs are buffer solution pool 1, buffer waste solution pool 2, sample pool 3 and sample injection waste solution pool 4. The sample pool 3 and the sample injection waste pool 4 are located on opposite sides of the separation channel. The overlapping area between the separation channel and the injection channel is the effective injection channel. All channels have a width of 150 μm and a depth of 50 μm. The effective sampling channel lengths are 0.5 cm, 10 cm and 20 cm, respectively, and the width is 150 μm. The inner surface of the chip channel is modified. The sample is Φx174 HaeIII Digest, the total DNA content is 2.5pg / μL, and there are 11 DNA fragments ranging from 72 to 1382 base pairs. The sample was dissolved in PCR reaction buffer containing 60mmol / L ...

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Abstract

The invention discloses a functionalized microfluidic chip and a method for analyzing PCR products. The chip is composed of a buffer pool, a buffer waste pool, a sampling waste pool, a sample pool, a separation channel and a sampling channel. , the two ends of the separation channel are respectively connected to the buffer pool and the buffer waste pool, the two ends of the sampling channel are respectively connected to the sample pool and the sampling waste pool, and the effective length of the sampling channel is 0.5-20 cm. The method of the invention combines isotachophoretic preconcentration and sieving electrophoresis separation, the buffer solution contains trailing dielectric medium and DNA sieving medium, and the self-contained chloride ion in the PCR product is used as the leading ion. The chip only needs to introduce a buffer solution, reduces the number of electrodes required, simplifies the operation, has the advantages of sensitivity and rapid analysis, and can effectively reduce the PCR reaction time by improving the detection sensitivity. The invention can analyze PCR products sensitively, rapidly and conveniently.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and in particular relates to a functionalized microfluidic chip for analyzing PCR products and a method for analyzing PCR products. Background technique [0002] Polymerase chain reaction (PCR) is one of the most widely used techniques in DNA research, through which the template DNA copy can be amplified by 10 6 times or more to meet analysis needs. The PCR product components are the amplified DNA fragments dissolved in the PCR buffer. [0003] The traditional PCR product analysis method is gel electrophoresis separation combined with gel imaging. The disadvantages of this method are low sensitivity and long amplification time for low-copy DNA templates. In addition, the method based on gel electrophoresis has the disadvantages of cumbersome operation and large sample consumption. In actual operation, a PCR product analysis method with high sensitivity, simple operation and fast an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12M1/34
Inventor 刘大渔
Owner GUANGZHOU IMPROVE MEDICAL TECH CO LTD
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