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A kind of microemulsion droplets of products after w/o/w type pcr amplification based on microfluidic system and preparation method thereof

A microfluidic system and post-product technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the waste of scientific research, the poor stability of W/O droplets, and the incomplete preservation of biological samples and other problems, to achieve the effect of small particle size change

Active Publication Date: 2022-06-03
长春维石检测技术服务有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It is well known that the stability of W / O droplets is relatively poor. Therefore, most amplified biological samples cannot be preserved intact for other experimental studies, which is undoubtedly a waste of scientific research.

Method used

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  • A kind of microemulsion droplets of products after w/o/w type pcr amplification based on microfluidic system and preparation method thereof
  • A kind of microemulsion droplets of products after w/o/w type pcr amplification based on microfluidic system and preparation method thereof
  • A kind of microemulsion droplets of products after w/o/w type pcr amplification based on microfluidic system and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

Embodiment 2

Embodiment 3

PCR system, using A3G‑VR1012 as the biological sample to design primers, as shown in Table 1; Table 2 is ddPCRSupermix for

Probes (no dUTP) PCR system; Table 3 is the ddPCR amplification conditions.

Table 1. is the primer and probe sequence of biological sample design with A3G-VR1012

[0046]

Table 2.ddPCR Supermix for Probes (no dUTP) PCR system

[0048]

[0049]

Table 3.ddPCR amplification conditions

[0051]

According to the ddPCR Supermix for Probes (no dUTP) PCR system of table 2, in Dolomite microfluidic system

The W / O microemulsion droplets packaged with the A3G‑VR1012 PCR system of different numbers of molecules were systematically prepared. Among them, pump 1 pumps different molecules

A3G‑VR1012, pump 2 pumped PCR system buffer solution, pump 3 pumped Droplet Generation Oil for

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Abstract

The invention provides a microemulsion droplet of a W / O / W type PCR amplified product based on a microfluidic system and a preparation method of the microemulsion droplet, the microemulsion droplet includes an amplified W / O type microemulsion droplet , an oil phase A and an outer water phase, wherein the amplified W / O microemulsion droplets include an inner water phase and an oil phase B, prepared by a microfluidic system, and the amplified W / O microemulsion droplets, oil phase A The volume ratio of the inner water phase to the outer water phase is 1:3:1000; the volume ratio of the inner water phase to the oil phase B is 40:3. The present invention provides a preparation method of W / O / W microemulsion droplets. The obtained microemulsion droplets can effectively establish a new type of microemulsion droplet sorting platform, and can effectively solve the problem of tissue The conundrum of cellular heterogeneity that samples cannot crack. In the present invention, by strictly screening the formula ratio and preparation parameters, the average particle diameter is 70 μm, all of which are spherical particles with uniform particle size distribution, and the stability experiment shows that the microemulsion droplets prepared by the present invention can be stored at room temperature, and The particle size changes little in a short time at room temperature.

Description

A kind of microemulsion droplets of products after W / O / W PCR amplification based on microfluidic system and the same Preparation technical field The invention belongs to biotechnology field, relate in particular to a kind of after W / O / W type PCR amplification based on microfluidic system Microemulsion droplets of products and methods for their preparation. Background technique With the establishment and development of single-cell genome sequencing and single-molecule sequencing technology, the analysis of cell genome characteristics The analysis went to the single-cell and single-molecule level. Single-cell genomic resolution not only enables researchers to Analyzing the heterogeneity of tumor cells also enables genomic studies of rare cells that have traditionally been difficult to detect. Depend on Since the size of cells is generally in the micrometer scale, microfluidic chip microdroplet technology has been widely used in cell sample processing. use. Cells...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2565/629C12Q2563/159
Inventor 王猛
Owner 长春维石检测技术服务有限公司
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