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Kit and method for extracting and purifying escherichia coli DNA from complicated sample

A technology of Escherichia coli and samples, applied in the field of molecular biology, can solve the problems of long time required and complicated process, and achieve the effect of saving experimental time and simplifying the experimental process.

Active Publication Date: 2018-03-23
山东凯景生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to make up for the deficiencies of the prior art and solve the problems of complicated process and long time required in the existing methods for extracting Escherichia coli DNA from complex samples, the present invention provides a kit for extracting and purifying Escherichia coli DNA from complex samples and method

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  • Kit and method for extracting and purifying escherichia coli DNA from complicated sample
  • Kit and method for extracting and purifying escherichia coli DNA from complicated sample
  • Kit and method for extracting and purifying escherichia coli DNA from complicated sample

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Embodiment 1

[0046] A kit for extracting and purifying Escherichia coli DNA from complex samples, characterized in that it comprises:

[0047] 1) Carboxylated nano-magnetic beads Eget103 coupled with gp37 protein;

[0048] 2) Sample diluent (reagent A);

[0049] 3) Sample Lysis Solution (Reagent B);

[0050] 4) Nucleic acid cleaning solution (reagent C);

[0051] 5) DNA Elution Solution (Reagent D)

[0052] 6) Nucleic acid adsorption magnetic beads;

[0053] in,

[0054] The composition of reagent A is: NaCl 137mM, KCl 2.7mM, NaCl 2 HPO 4 4.3mM, KH 2 PO 4 1.4mM, 0.05% Tween-20.

[0055] The composition of reagent B is: NaCl 0.05mM, EDTA 5mM, Tris-HCl 20mM with pH=8.0, lysozyme 3mg / ml.

[0056] The composition of reagent C is: NaCl 0.05mM, EDTA 0.05mM, Tris-HCl 20mM at pH=8.0, absolute ethanol 70%.

[0057] The composition of reagent D is: EDTA 0.05mM, Tris-HCl 10mM at pH=8.0.

[0058] The preparation method of carboxyl nano magnetic beads Eget103 coupled with gp37 protein is ...

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Abstract

The invention discloses a kit and a method for extracting and purifying escherichia coli DNA from a complicated sample, and belongs to the field of molecular biology. The kit for extracting and purifying the escherichia coli DNA from the complicated sample comprises (1) carboxylated nano magnetic bead Eget103 coupled with gp37 protein, (2) sample diluent, (3) sample lysate, (4) nucleic acid cleaning fluid, (5) DNA eluent, and (6) nucleic acid adsorption magnetic beads. According to the kit and the method, the carboxylated nano magnetic bead Eget103 coupled with gp37 protein are specifically combined with escherichia coli to purify the escherichia coli, and then DNA is extracted from the purified escherichia coli by a magnetic bead method. In the whole process, no isolation and purificationculture of the escherichia coli is needed, and the DNA is directly extracted; therefore, the experimental process is simplified, and the experimental time is greatly shortened.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a kit and method for extracting and purifying Escherichia coli DNA from complex samples. Background technique [0002] Escherichia coli, also known as Escherichia coli (Gram-negative short bacilli), is a common resident bacteria in the intestinal tract of humans and animals. According to different pathogenicity, diarrhea-causing Escherichia coli is divided into enterotoxigenic Escherichia coli, enteroinvasive Escherichia coli, enteropathogenic Escherichia coli, enteroaggregative adhesive Escherichia coli and enterohaemorrhagic Escherichia coli 5 species. [0003] T4 bacteriophage is a type of virus that takes Escherichia coli as a host, and has strong host specificity. The gp37 protein is located at the end of the long tail filament of T4-like phages, and its C-terminus can recognize the surface receptors of host bacteria, triggering the process of phage adsorption to the surfa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2523/308C12Q2527/125
Inventor 李硕张通王义翠刘奕
Owner 山东凯景生物技术有限公司
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