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PDMS micro-control board treatment method and application thereof

A treatment method and micro-control technology, applied in the field of biological experiments, can solve the problems of unstable properties of BSA, unstable test results, and high price of BSA, so as to reduce cell adhesion, maintain cell integrity, and improve test accuracy. Effect

Pending Publication Date: 2019-04-05
SINGLERON NANJING BIOTECHNOLOGIES LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of the prior art are: 1. BSA itself is unstable and needs to be stored at 2-8°C under harsh storage conditions, and it needs to be prepared and used immediately, and it is not resistant to storage
2. BSA contains a small amount of nuclease, which may degrade RNA, and the difference between different batches is large, resulting in unstable test results
3. The price of BSA is high and the cost of experiment is high

Method used

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  • PDMS micro-control board treatment method and application thereof
  • PDMS micro-control board treatment method and application thereof
  • PDMS micro-control board treatment method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Prepare 0.02% PBST solution: prepare 20% Tween-20 with nuclease free water, then prepare 0.02% PBST according to the ratio of PBS:20% Tween-20=1000:1, vortex and mix well.

[0031] S1: PDMS surface treatment: use a pipette to absorb 200 μL of 0.02% PBST solution, add it to the surface of PDMS, and vacuum it with a vacuum desiccator for 15 minutes.

[0032] S2: Add cells: Add 100 μL of cell suspension (resuspended in PBS), let stand for 2 minutes, let the cells fall into the microwell, and then use a pipette to absorb 500 μL of PBS solution to wash away the excess cells remaining on the surface. The cells used in the present invention are mouse embryonic fibroblasts L929.

[0033] S3: Capturing mRNA: Add 100-100 μL of cell lysate to lyse the cells to obtain mRNA. Aspirate the solution with a pipette.

[0034] S4: Reverse transcription and PCR amplification: Reverse transcription and PCR amplification were performed using Novizyme’s reverse transcription reagent (Cat. N...

Embodiment 2

[0048] Other conditions remained unchanged, the PDMS surface was treated with 0.01% PBST for 10 min, and the cells used were A431. DNA fragment detection is shown in Table G-I below.

Embodiment 3

[0050] Other conditions remained unchanged, the PDMS surface was treated with 0.1% PBST for 10 minutes, and the cells used were 293T. For DNA fragment detection, see Tables J-L below.

[0051] The present invention has been verified by samples, which proves the feasibility of the method. Now list the results of a comparative experiment we have done. We use conventional PBS+1%BSA solution and 0.02%PBST solution to treat PDMS. more stable. In the process of several experiments, the existing technology has great variability, but the technical solution has less variability and more stable performance, which is conducive to improving the accuracy of the experiment.

[0052]

[0053]

[0054] Figure 1-4These are the purified products of C, F, I, and L above, and the size of the DNA fragment was detected with the FragmentAnalyzer of the AATI automatic capillary electrophoresis instrument. The surface of PDMS can reduce the adhesion of cells without harming the cells and ma...

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Abstract

The invention relates to a biological experiment method, in particular to a PDMS micro-control board treatment method and application thereof. The PDMS micro-control board treatment method comprises the following steps that S1, PDMS surface treatment is carried out, specifically, a PBST solution is absorbed, the PBST solution is added to the PDMS surface, and the treatment time is carried out for10-15min; S2, cells are added, specifically, a cell suspension is added, standing is carried out, the mixture is washed with PBS to wash away excess cells left on the surface; S3, mRNA is captured, specifically, cell lysate is added to lyse the cells to obtain the mRNA; S4, reverse transcription and PCR amplification are carried out, specifically, reverse transcription is carried out on the captured mRNA to obtain cDNA, and PCR amplification is carried out on the cDNA to obtain PCR products; and S5: DNA is purified. The treatment method can reduce the surface adsorbability and increase the compatibility of PDMS by treating the surface, so that the test accuracy is improved; and the method has the advantages of simple and convenient operation, good effect, low cost and the like.

Description

technical field [0001] The invention relates to a biological experiment method, in particular to a PDMS micro-control board processing method and application thereof. Background technique [0002] PDMS (polydimethylsiloxane, polydimethylsiloxane), is a polymer organosilicon compound, solid PDMS is a kind of silicone, hydrophobic (hydrophobic) and waterproof, non-toxic, non-flammable, transparent elastomer . Because of its simple and fast production, low cost, good adhesion to silicon wafers, and good chemical inertness, it has become an organic polymer material widely used in microfluidics and other fields, especially common Biological microplate microchip material. Due to the hydrophobicity of PDMS, it is incompatible in biological experiments, and its surface needs to be treated to make it compatible. At present, most experiments pre-treat the PDMS surface by PBS (phosphate buffer saline, phosphate buffered saline) + 1% BSA (Albumin from bovine serum, bovine serum album...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1006
Inventor 吴双方南周璟
Owner SINGLERON NANJING BIOTECHNOLOGIES LTD