Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Screening method of gene polymorphism detection probe

A gene polymorphism and detection probe technology, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., to achieve the effect of saving research and development costs and saving research and development time

Active Publication Date: 2018-08-03
深圳会众生物技术有限公司
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a screening method for gene polymorphism detection probes based on the deficiencies of the above-mentioned detection schemes, which can avoid the problems of high cost and low research and development efficiency caused by the screening of a large number of probes

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Screening method of gene polymorphism detection probe
  • Screening method of gene polymorphism detection probe
  • Screening method of gene polymorphism detection probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035]Taking the screening of the gene polymorphism detection probe of folate metabolism gene MTHFR as an example, the screening method of the gene polymorphism detection probe of the present invention is used to screen the MTHFR gene polymorphism detection probe, and its complete technical scheme includes:

[0036] (1) Preparation and concentration determination of sample genomic DNA

[0037] Source of samples: human blood samples, oral cells and amniotic fluid; Genomic DNA was prepared using Micro Sample Genomic DNA Extraction Kit (spin column type), and the extracted DNA was measured with Nanodrop2000 to determine the concentration and purity of the extracted DNA, and then diluted to a certain Subsequent amplification verification work was carried out in the concentration range.

[0038] (2) Design of probes and primers

[0039] The polypeptide sites of the MTHFR gene are C677T and A1298C. According to the base sequence of each mutation site of MTHFR in the GeneBank databa...

Embodiment 2

[0057] Taking the screening of the gene polymorphism detection probe of folic acid metabolism gene MTRR as an example, the screening method of the gene polymorphism detection probe of the present invention is used to screen the MTRR gene polymorphism detection probe, and its complete technical scheme includes:

[0058] (1) Preparation and concentration determination of sample genomic DNA

[0059] Source of samples: human blood samples, oral cells and amniotic fluid; Genomic DNA was prepared using Micro Sample Genomic DNA Extraction Kit (spin column type), and the extracted DNA was measured with Nanodrop2000 to determine the concentration and purity of the extracted DNA, and then diluted to a certain Subsequent amplification verification work was carried out in the concentration range.

[0060] (2) Design of probes and primers

[0061] The polypeptide site of the MTRR gene is A66G. According to the base sequence of the mutation site in the GeneBank database, specific PCR ampli...

Embodiment 3

[0079] This example is basically the same as Example 1, except that this example further discloses gene templates and universal probe sequences involved in the specific screening process. For the MTHFR gene templates and universal probe sequences (10 each) used in the screening process of this example, see Table 5 below for details.

[0080] table 5

[0081]

[0082]

[0083] Table 6 is a list of preferred MTHFR templates, primers and universal probe sequences obtained after screening.

[0084] Table 6

[0085]

[0086]

[0087] The " / " in Table 6 indicates the specific separation position between the first universal probe sequence and the second universal probe sequence, the specific separation position between the first template sequence and the second template sequence, the first universal probe sequence and the second universal probe sequence formed by sequential separation. A specific separation position between a primer sequence and a second primer sequence...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a screening method of a gene polymorphism detection probe. The method comprises the following steps of (1) designing a universal probe according to a bacterium 16S rDNA conserved region; designing a template sequence containing a mutation site; (2) designing a plurality of pairs of screening primers P1 and a plurality of screening primers P2 covering polymorphism sites according to the base sequence of the mutation site of a gene to be tested of a GeneBank database; (3) gradually taking one pair of primers in the plurality of pairs of screening primers P1 and the plurality of screening primers P2 obtained and designed in the step (2) one by one to perform annealing reaction with the universal probe and the template sequence designed and obtained in the step (1); performing screening to obtain the preferable template sequence; (4) according to the preferable template sequence, synthesizing the matched fluorescent probe, i.e., the final preferable probe. The screening method of the gene polymorphism detection probe can avoid the problems of high cost and low research and development efficiency caused by screening of a great number of probes can be avoided.

Description

technical field [0001] The invention relates to gene detection technology, in particular to a method for screening gene polymorphism detection probes. Background technique [0002] Existing gene detection technologies include: (1) Sanger sequencing: it is the gold standard for detecting gene mutations, but due to its time-consuming, cumbersome operation, high cost, and limited clinical application; (2) PCR-fluorescence melting curve method: using A method for detecting gene mutations with different double-stranded DNA melting Tm values. It usually includes dye method and probe method. When this method is used for multiple asymmetric amplification and multi-site detection, the amplification difficulty increases, and the difference in Tm value is difficult to distinguish. Multi-tube amplification is often required, which reduces the detection throughput. Higher cost; (3) Fluorescent PCR method: use fluorescently labeled probes to specifically bind to target sequences of diffe...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/6811
CPCC12Q1/6811C12Q2563/107C12Q2545/101
Inventor 刘晶晶
Owner 深圳会众生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com