CRISPR assisted DNA target enrichment method and application thereof

A DNA targeting and DNA library technology, applied in the field of biomedicine, can solve the problem of reducing the specificity of target enrichment and achieve low-cost effects

Active Publication Date: 2019-06-04
SOUTHEAST UNIV
View PDF4 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Additionally, non-specific hybridization is unavoidable and the resulting hi

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
  • CRISPR assisted DNA target enrichment method and application thereof
  • CRISPR assisted DNA target enrichment method and application thereof
  • CRISPR assisted DNA target enrichment method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Utilize CATE technology to enrich 35 exon sequences of 6 genes in the genomic DNA of 6 cell lines, and use cloning sequencing and next-generation sequencing (NGS) sequencing analysis; Utilize CATE technology to enrich the mutant TERT promoter sequence and use ARMS-qPCR.

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 discloses a CRISPR assisted DNA target enrichment method and an application thereof. sgRNA of a CRISPR/Cas9 system is reformed, novel sgRNA having an acquisition sequence at a 3' terminal is developed, the sgRNA and Cas9 protein without nuclease activity form a compound, the compound and target DNA are in target combination, and a formed DNA-dCas9-sgRNA compound can be acquired by magnetic beads of which the surfaces are fixed with single-chain acquired oligonucleotide, so that the target DNA is in target enrichment and separation from a DNA library or a mixture, and the CRISPR assisted DNA target enrichment method is used for sequence analysis of the target DNA. Compared with a target enrichment method based on hybridization, which is universal at present, the method provided by the invention has notable advantages of being high in simplicity, high in specificity, high in sensitivity, high in flux and the like, and can be deeply applied to preparation of reagents for DNAdetection, diagnosis and treatment.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a CRISPR-assisted DNA targeted enrichment sequencing method and its application. Background technique [0002] Current medicine, especially personalized or precision medicine, increasingly relies on DNA analysis. DNA from clinical samples is increasingly used to find diagnostic, prognostic, and predictive biomarkers of disease. DNA analysis also opens up new opportunities for disease diagnosis through liquid biopsy, in vitro diagnostics (IVD) and non-invasive prenatal testing (NIPT). DNA analysis can be used to discover genetic and epigenetic alterations occurring in the genome, such as single nucleotide variants (SNVs), copy number variants (CNVs), translocations and methylation. With the widespread application of DNA sequencing technology, next-generation sequencing (NGS) provides a powerful tool capable of decoding DNA-encoded information at a genome-wide sca...

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): C12N15/10C12Q1/6806
Inventor 王进科徐新慧
Owner SOUTHEAST 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