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

Method for fast identification of recombinant agrobacterium tumefacien plasmids

An identification method, the technology of Agrobacterium, which is applied in the field of scientific research, can solve the problems of complex and cumbersome operation process and the hazards of operators, and achieve the effects of simplifying the operation process, reducing the harm, and improving the extraction volume and purity

Inactive Publication Date: 2012-10-17
JILIN NORMAL UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Based on the particularity of Agrobacterium cells, it is generally necessary to add a certain amount of lyase during extraction, and after extraction, it needs to be purified with phenol, chloroform, etc. have direct or indirect harm

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0024] The material that the inventive method adopts comprises:

[0025] (1) Strains: Agrobacterium rhizogenes A4 strain, 15834 strain (preserved by the Laboratory of Genetic Physiology Institute of Jilin Normal University), Agrobacterium tumefaciens GV3101 strain (preserved by the Laboratory of Genetic Physiology Institute of Jilin Normal University);

[0026] (2) Plasmid: pCAMBIA1302 (kan resistance) (purchased from pCAMBIA company in Australia).

[0027] The rapid identification method of the recombinant Agrobacterium plasmid comprises the following steps:

[0028] (1) After transforming the plasmid pCAMBIA1302 into the A4 strain or 15834 strain of Agrobacterium rhizogenes or the GV3101 strain of Agrobacterium tumefaciens; through the method of coating culture, in the ultra-clean workbench, spread the transformed bacterial solution on the carrying On the petri dish of kan-resistant solid LB medium, seal the petri dish with a sealing film to ensure aseptic conditions, and p...

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 for identification of agrobacterium tumefaciens and especially relates to a method for fast identification of recombinant agrobacterium tumefacien plasmids. The method can be directly used in the field of a scientific research. The method comprises the following steps of transferring plasmids to agrobacterium tumefaciens by a heat shock method or an electroporation method, carrying out a spreading culture by a solid medium with antibiotics to obtain a single colony, picking the single colony, carrying out a shake-flask culture until OD600 is in a range of 1.0 to 1.2, carrying out centrifugation, collecting thalluses, adding a suspension into the thalluses, mixing uniformly, adding lysozyme into the mixture, boiling for 1 to 3 minutes, carrying out centrifugation, separating recombinant plasmids from empty carriers by an agarose gel electrophoresis, carrying out polymerase chain reaction (PCR) detection, and identifying positive clones.

Description

technical field [0001] The invention relates to an identification method of Agrobacterium, in particular to a rapid identification method of recombinant Agrobacterium plasmid, which can be directly applied to the field of scientific research. Background technique [0002] Plasmid extraction and purity identification of recombinant Agrobacterium is a routine experimental operation in molecular cloning and Agrobacterium-mediated plant genetic transformation research, and whether the plasmid is successfully transformed has an important impact on subsequent experiments. At present, the conventional methods for extracting Agrobacterium plasmid DNA mainly include alkaline lysis, Mini plasmid extraction, boiling and the like. Based on the particularity of Agrobacterium cells, it is generally necessary to add a certain amount of lyase during extraction, and after extraction, it needs to be purified with phenol, chloroform, etc. There are direct or indirect hazards. Contents of 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 Applications(China)
IPC IPC(8): C12Q1/68C12Q1/02
Inventor 周晓馥未晓巍勾畅
Owner JILIN NORMAL UNIV
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