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

Method, composition and kit for performing in-vitro nucleic acid unidirectional amplification by utilizing transposon enzyme

A technology of transposase and transposase, which is applied in the field of molecular biology, can solve problems such as inability to capture, and achieve the effect of overcoming false positives

Active Publication Date: 2016-12-14
GENEIS TECH BEIJING CO LTD
View PDF5 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fragmentation of genomic DNA can be achieved using such technologies, but this technology still cannot capture specific sequences on a genome-wide scale

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, composition and kit for performing in-vitro nucleic acid unidirectional amplification by utilizing transposon enzyme
  • Method, composition and kit for performing in-vitro nucleic acid unidirectional amplification by utilizing transposon enzyme
  • Method, composition and kit for performing in-vitro nucleic acid unidirectional amplification by utilizing transposon enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0051] Unless otherwise specified, any reagent used in the present invention is a commercially available general reagent. The blood samples were collected from the Union Hospital.

[0052] 1. Sample extraction

[0053] Blood DNA was extracted using DNA extraction reagents. The main steps are as follows: temporarily store the whole blood sample in a refrigerator at 4°C. The blood DNA extraction kit from QIAgene Company of the United States was used for extraction. For specific steps, please refer to the instruction manual attached to the kit, and details will not be repeated here. Quantitative analysis was performed on the extracted DNA to ensure that each extracted DNA exceeded 200ng. Thus, 10 copies of DNA were obtained.

[0054] 2. Transposonase reaction

[0055] Thaw 5×Tagment buffer L at room temperature, invert up and down and mix well before use; confirm that 5XNT Solution is at room temperature, and flick the tube wall to confirm that there is no precipitation.

...

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, a composition and a kit for performing in-vitro nucleic acid unidirectional amplification by utilizing a transposon enzyme. According to the method, the composition and the kit, a target area amplifying process is completed by combining the transposon enzyme and unidirectional specific amplification and high-throughput sequencing methods, and the original DNA mutation frequency is restored very well. The detection of a specific DNA sequence can be realized, and the purpose of detecting tumour cell mutations or genetic disease genes can be achieved. The problems that hybridization is required by a conventional method or the bias exists in massive PCR amplification are solved; meanwhile, the mutation frequency is reserved, and the problems of a false positive of a sample caused by a sequencing error and an amplification error are solved.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a method, composition and kit for unidirectional amplification of nucleic acid molecules by using transposonase. Background technique [0002] There are three billion base pairs in the human genome. Selecting some bases, such as the pathogenic part, for analysis is the main clinical need at present. How to capture specific sequences in the whole genome is a world-wide challenge. sexual problem. [0003] Transposon enzyme is an enzyme that performs the transposition function, usually encoded by a transposon, which recognizes the specific sequence at both ends of the transposon, and can detach the transposon from the adjacent sequence and insert it into the new DNA Target site, no homology requirement. At present, transposon enzymes have been modified to allow transposon enzymes to insert specific sequences (for example, NEXTERA from Epicentre TM system). Genomic DNA can be fra...

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): C12N15/10
CPCC12P19/34C12Q1/6806C12Q2523/109C12Q2531/113
Inventor 田埂
Owner GENEIS TECH BEIJING CO LTD
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