Method utilizing fluorescence PCR technology to identify mouse genotype

A type identification, mouse technology, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of cumbersome, false negative, low resolution, etc.

Inactive Publication Date: 2015-12-23
DONGHUA UNIV
View PDF4 Cites 36 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is too cumbersome and needs to be annealed for a period of time before the next experiment can be carried out; using the enzyme diges...

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
  • Method utilizing fluorescence PCR technology to identify mouse genotype
  • Method utilizing fluorescence PCR technology to identify mouse genotype
  • Method utilizing fluorescence PCR technology to identify mouse genotype

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Mouse feeding and sample collection: The identified Founder mice (I used the Crispr / Cas9 system to knock out mircoRNA505 (GeneID: 751545) C57BL / 6J mice were used as Founder mice, and the Founder mice were identified Knockout 23bp (denoted as Founder17) was crossed with C57BL / 6J mice to generate F1 generation mice. Take 1 cm tail tissue from F1 generation mice and store at -20°C for future use.

[0035] (2) DNA extraction: Use the animal genome DNA extraction kit from Sangon Bioengineering Co., Ltd. to extract (1) the middle tail tissue according to the instructions to obtain DNA, and determine the DNA quality and concentration by 0.8% agarose gel electrophoresis .

[0036](3) Primer design: Design a pair of PCR primers in the region of the gene locus knocked out by the Crispr / Cas9 system, and use Primer3 online software (http: / / frodo.wi .mit.edu / primer3 / ) for preliminary primer design, then artificially designed with oligo6, and synthesized by Shanghai Sangon Bioe...

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 method utilizing a fluorescence PCR technology to identify mouse genotype. A fluorescence PCR primer is utilized, after PCR amplification, the amplification product is mixed with ROX with a known size, the size of PCR products can be calculated by Genemapper software through 377 sequencing; and thus the mouse genotype can be identified. The provided method can simply, directly, and precisely find out the mice having micro RNA, whose Crispr/Cas9 has been knocked off.

Description

technical field [0001] The invention belongs to the field of mouse typing identification methods, in particular to a mouse typing identification method using fluorescent PCR technology. Background technique [0002] Since its inception, gene targeting technology has been one of the important means to study gene function, revealing the biological functions of many important genes. In addition, researchers also hope to use gene targeting technology to knock out or modify specific genes, so as to achieve the purpose of treating diseases or improving production traits of livestock and poultry. However, the efficiency of early gene targeting technology was extremely low, and it was difficult to truly apply it to medical treatment or livestock and poultry improvement. CRISPR / Cas9 system, as an emerging gene-directed editing technology, has gradually matured and been successfully applied in many animal and plant species, which has greatly promoted the study of gene function. The ...

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): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2600/158C12Q2600/178C12Q2545/101C12Q2563/107C12Q2565/125
Inventor 肖君华李雨王茂春仝莉周宇荀
Owner DONGHUA UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap