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Functionalization micro-flow control chip and 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, etc., and achieve the effect of increasing the injection volume, fast and convenient PCR product analysis, and simplifying the operation

Active Publication Date: 2008-04-09
GUANGZHOU IMPROVE MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Functionalization micro-flow control chip and method for PCR product analysis
  • Functionalization micro-flow control chip and method for PCR product analysis
  • Functionalization micro-flow control chip and method for PCR product analysis

Examples

Experimental program
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Effect test

Embodiment 1

[0039] A functionalized microfluidic chip made of PMMA is used. The configuration is shown in Figure 3. The chip consists of 4 reservoirs, a separation channel and a sampling channel. The 4 reservoirs are buffer pool 1, buffer waste Liquid pool 2, sample pool 3 and sampling waste 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 2.5pg / μL, and there are 11 DNA fragments r...

Embodiment 2

[0046] The configuration of the functionalized microfluidic chip made of glass is shown in Figure 5. The chip consists of 4 reservoirs, a separation channel and a sampling channel. The 4 reservoirs are buffer pool 1, buffer waste pool 2, sample pool 3 and sample injection waste 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 as the trailing dielectric, 40mmol / L imidazole, 1 μmol / L S...

Embodiment 3

[0053] The functionalized microfluidic chip made of PDMS has a configuration as shown in Figure 3. The chip consists of 4 reservoirs, a separation channel and a sampling channel. The 4 reservoirs are buffer pool 1, buffer waste Liquid pool 2, sample pool 3 and sampling waste 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 chloride ion. The composition of the separation buffer i...

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Abstract

The invention discloses a functionalized microflow controlled chip and a method using the functionalized microflow controlled chip to analysis the PCR products. The chip comprises a buffering liquid pool, a waste liquid pool of buffering liquid, a sample injection waste liquid pool, a sample pool, a separating channel and a sample injection channel. Two ends of the separating channel are respectively connected with the buffering liquid pool and the waste liquid pool of buffering liquid. Two ends of the sample injection channel are respectively connected with the sample pool and the sample injection waste liquid pool. The length of the effective sample injection channel is 0.5cm to 20cm. the invention combines the isotachophoresis pre concentration and the screening electrophoresis separation. The buffering liquid contains a liac dielectric medium and a DNA screening medium. The method uses the chloride ion self contained in PCR products as a leading ion. The chip is only needed to be introduced in one type of buffering liquid, which reduces the number of the needed electrode and simplify the operation and has the advantages of sensitive and fast analyzing. The chip can effectively shorten the PCR reaction time through enhancing the detecting sensitivity. The invention can sensitively, fast and simply analyze the PCR products.

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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IPC IPC(8): C12Q1/68C12M1/34
Inventor 刘大渔
Owner GUANGZHOU IMPROVE MEDICAL TECH CO LTD
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