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Kit and device for ATRX and KDM5A mutation detection based on digital PCR technology and application

A mutation detection and kit technology, applied in the field of sequencing libraries, can solve problems such as low accuracy and poor detection efficiency of kits, and achieve high specific surface area, enhanced mixed reaction effect, and precise effect

Pending Publication Date: 2022-07-12
北京组学生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing kits based on digital PCR technology have poor detection efficiency and low accuracy. Based on this, further improvement is needed

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0023] The preparation method of described sizing agent is:

[0024] Mix 5-10 parts of sodium lauryl sulfate, 1-3 parts of hydrochloric acid and 1-5 parts of chitosan solution, the mixing speed is 100-300r / min, the mixing time is 10-20min, and the mixing is completed;

[0025] Then add 1-5 parts of modified silica, continue to stir at a speed of 100-200rmin for 20-30min, the stirring is completed, and then add 1-3 parts of modified wollastonite to obtain a sizing agent.

[0026] The modification method of the modified silica in this embodiment is as follows: the silica is sent to 100-300°C for calcination for 10-20min, the calcination is finished, then the temperature is changed to 50-60°C, and then sent to water for static Set aside and cool to room temperature.

[0027] The specific operation steps of the temperature-change-down treatment in this embodiment are: dropping to 50-60°C at a rate of 1-5°C / min.

[0028] The mass fraction of the chitosan solution in this embodime...

Embodiment 1

[0035] The kit for detecting ATRX and KDM5A mutations based on digital PCR technology in this embodiment includes mutant probes, infiltrating agents and additives in a weight ratio of 4:2:1;

[0036] The preparation method of described sizing agent is:

[0037] Mix 5 parts of sodium lauryl sulfate, 1 part of hydrochloric acid and 1 part of chitosan solution, the mixing speed is 100r / min, the mixing time is 10min, and the mixing is completed;

[0038] Then, 1 part of modified silica was added, and the stirring was continued for 20 min at a rotating speed of 100 rmin, and the stirring was completed, and then 1 part of modified wollastonite was added to obtain a sizing agent.

[0039] The modification method of the modified silica in this embodiment is as follows: the silica is sent to 100°C for calcination for 10min, the calcination is completed, then the temperature is changed to 50°C, then put into water and left to stand, and cooled to room temperature .

[0040] The specif...

Embodiment 2

[0047] The kit for detecting ATRX and KDM5A mutation based on digital PCR technology in this embodiment includes mutant probe, infiltrating agent and additives in a weight ratio of 4:2:1;

[0048] The preparation method of described sizing agent is:

[0049] Mix 10 parts of sodium lauryl sulfate, 3 parts of hydrochloric acid and 5 parts of chitosan solution, the mixing speed is 300r / min, the mixing time is 20min, and the mixing ends;

[0050] Then, 5 parts of modified silica were added, and the stirring was continued for 30 minutes at a rotating speed of 200 rmin, and the stirring was completed, and then 3 parts of modified wollastonite were added to obtain a sizing agent.

[0051] The modification method of the modified silica in this example is as follows: the silica is sent to 300°C for calcination for 20min, the calcination is completed, then the temperature is changed to 60°C, then put into water and left to stand, and cooled to room temperature.

[0052] The specific op...

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PUM

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Abstract

The invention provides an ATRX and KDM5A mutation detection kit based on a digital PCR technology. The ATRX and KDM5A mutation detection kit comprises a mutant probe, an impregnating compound and an additive according to a weight ratio of 4: 2: 1, the preparation method of the impregnating compound is as follows. The kit is prepared from the mutant probe, the impregnating compound and the additive, the mutant probe is used for detecting genes, a chitosan solution and lauryl sodium sulfate in the impregnating compound can enable the probe to better detect the genes, the effect is more accurate, and meanwhile, due to the high specific surface area of modified silicon dioxide, the detection sensitivity is improved. The mixing reaction effect of the product raw materials is enhanced; and the added additive improves the capability of the raw materials, so that the detection accuracy is further improved.

Description

technical field [0001] The invention relates to the technical field of sequencing libraries, in particular to a kit, device and application for detecting ATRX and KDM5A mutations based on digital PCR technology. Background technique [0002] Next-generation sequencing is also called deep sequencing and massively parallel sequencing. [0003] At present, there are mainly three mainstream sequencing platforms: Hiseq platform of illumina company; high-throughput sequencing technology is a revolutionary change to traditional sequencing, which can sequence hundreds of thousands to millions of DNA molecules at a time. In some literatures, it is called next generation sequencing, which shows its epoch-making changes. At the same time, high-throughput sequencing makes it possible to conduct detailed and comprehensive analysis of the transcriptome and genome of a species, so it is also called deep sequencing. . [0004] Existing kits based on digital PCR technology have poor detect...

Claims

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Application Information

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IPC IPC(8): C12Q1/686C12N15/11C40B50/06C12Q1/6806C12Q1/6876C12M1/28C12M1/34C12M1/00
CPCC12Q1/686C40B50/06C12Q1/6806C12Q1/6876C12Q2600/156C12Q2527/125C12Q2535/122
Inventor 周小龙黄莉莎刘连成王运斌
Owner 北京组学生物科技有限公司