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

A method and kit for enriching circulating tumor DNA based on multiplex PCR

A kit and tumor technology, applied in the direction of tumor/cancer cell, biochemical equipment and method, microorganism determination/inspection, etc., can solve the problems of sample loss, sensitivity reduction, etc., to reduce errors, reduce false positives, and reduce effect of loss

Active Publication Date: 2021-03-23
常州桐树生物科技有限公司
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the current detection processes will have a magnetic bead purification step in the middle, and each purification step will inevitably lead to sample loss, resulting in a decrease in detection sensitivity

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
  • A method and kit for enriching circulating tumor DNA based on multiplex PCR
  • A method and kit for enriching circulating tumor DNA based on multiplex PCR
  • A method and kit for enriching circulating tumor DNA based on multiplex PCR

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] The method of the invention is used to detect ctDNA tumor mutations in blood samples.

[0097] Specific steps are as follows:

[0098] 1. Test template preparation.

[0099] The cultured SK cell line and HCC1975 (containing EGFR: p.L858R mutation) cell line were subjected to DNA extraction according to the phenol-chloroform method, and then DNA fragmentation was performed with fragmentation enzyme from NEB Company. Then the DNA of the HCC1975 sample was incorporated into the SK sample at a ratio of 1%. The EGFR:p.L858R mutation frequency was quantified by ddPCR to obtain samples containing the EGFR:p.L858R mutation with known mutation frequency.

[0100] 2. Primer design. This embodiment designs 2 kinds of PCR primers

[0101] Primer I: PCR primer without U base.

[0102] Primer II: PCR primer with U base.

[0103] The length of the designed primers is 60-70bp, the formed primer-dimer sequence is between 70-100, the length of the designed universal sequencing adap...

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 method and kit for enriching circulating tumor DNA on the basis of multiplex PCR. The method comprises the steps of (1), designing multiple PCR primer pairs, wherein PCR primers comprise an amplifying primer sequence used for amplifying a gene target region and a primer ligation sequence, and the primer ligation sequence is used for being linked with a universal sequencing linker of a sequencing platform; (2), taking a template containing the circulating tumor DNA and making the template and the PCR primer pairs in step (1) subjected to a first PCR reaction to obtaina first PCR reaction product; (3), adding a combined enzyme into the first PCR reaction product to digest residual PCR primers; (4), adding the universal sequencing linker into the first PCR reactionproduct and carrying out a second PCR reaction to obtain a sequencing library. The method and the kit have the effect of reducing sample loss.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method and kit for enriching circulating tumor DNA based on multiplex PCR. Background technique [0002] With the completion of the human genome completion map, human beings have entered the post-genome era. In the post-genome era, it is first required to use the DNA sequence information of the human genome to study genes and their variations in order to detect the functional changes brought about by the variations. [0003] Usually, the occurrence and development of tumors involves the occurrence and development of multiple genes, and the research on the change of a single gene has shown great limitations. Only by comprehensively detecting the base mutations of multiple genes can effective monitoring of MRD, medication guidance, therapeutic efficacy and surgical procedures be carried out. Post-treatment etc. [0004] In recent years, extensive research has been carried out on th...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09C12Q1/686
CPCC12N5/0693C12Q1/686C12Q2537/143
Inventor 蔡微菁徐泽
Owner 常州桐树生物科技有限公司
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