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Kit and method for extracting and purifying Salmonella DNA from complex samples

A Salmonella, sample-based technology, applied in the field of molecular biology, can solve problems such as complex process and long time required, and achieve the effect of simplifying the experimental process and saving experimental time

Active Publication Date: 2021-04-06
JINAN KAICHEN BIOTEC
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  • Summary
  • 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 Salmonella DNA from complex samples, the present invention provides a kit and method for extracting and purifying Salmonella DNA from complex samples

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  • Kit and method for extracting and purifying Salmonella DNA from complex samples
  • Kit and method for extracting and purifying Salmonella DNA from complex samples
  • Kit and method for extracting and purifying Salmonella DNA from complex samples

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Embodiment 1

[0046]A kit extracted and purified from Salmonella DNA from complex samples, including:

[0047]1) CPBA-MNPS CPBA-MNPs in the carboxygenic nanomagnetic beads of GP13 protein;

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

[0049]3) Sample cracking solution (reagent B);

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

[0051]5) DNA eluent (reagent D);

[0052]6) Nucleic acid adsorption beads;

[0053]among them,

[0054]The composition of the reagent A is: NaCl 137mm, KCl 2.7mm, Na2HPO4 4.3mm, kh2PO4 1.4mm, 0.05% Tween-20.

[0055]The composition B was formulated: NaCl 0.05 mm, EDTA 5 mM, pH = 7.5 TRIS-HCl 20 mM, lysozyme 3 mg / ml.

[0056]The composition C was completed as: NaCl 0.05 mm, EDTA 0.05 mM, pH = 7.5 TRIS-HCl 20 mM, and 70% absolute ethanol.

[0057]The composition of the reagent D was: EDTA 5 mM, pH = 8.8 Tris-HCl 10 mm.

[0058]The preparation method of carboxy nanomagnetic beads CPBA-MNPs in GP13 protein is:

[0059]a) Salmonella expression system using GP13 protein, soluble expression GP13 protein and purifi...

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Abstract

The invention discloses a kit and a method for extracting and purifying Salmonella DNA from complex samples, belonging to the field of molecular biology. The kit for extracting and purifying Salmonella DNA from complex samples of the present invention includes: 1) carboxylated nano magnetic beads CPBA-MNPs coupled with gp13 protein; 2) sample diluent; 3) sample lysate; 4) nucleic acid cleaning 5) DNA eluent 6) Nucleic acid adsorption magnetic beads. The present invention uses carboxylated nano-magnetic beads CPBA-MNPs coupled with gp13 protein to specifically bind Salmonella and then purify Salmonella, and then extract DNA from the purified Salmonella by using a magnetic bead method. The whole process does not require the isolation and purification of Salmonella, and DNA extraction is performed directly; the experimental process is simplified and the experimental time is greatly saved.

Description

Technical field[0001]The present invention relates to the field of molecular biology, and in particular, to a kit and method for extracting and purifying Salmonella DNA from complex samples.Background technique[0002]salmonella(Salmonella ) Is Gram-negative intestinal bacteria, a common human beast common disease, not only causes various animal diseases, but also related to human beings. Salmonella can be divided into 2 species of intestinal Salmonella and Banggol Salmonella, and the intestinal salmonella can be divided into 6 subspecies.[0003]Salmonella P22 is a class of viruses with Salmonella, and the host is very specific. The GP13 is located in the phage P22 long tail filament end, and the C-terminal can identify the surface receptor of the host bacteria, which initiates the adsorption of the phage to the surface of the Salmonella, and is a key protein of the phage P22 recognition receptor.[0004]Complex samples refer to food, feces, soil, vomit, etc., which typically contain inh...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10C12R1/42
CPCC12N15/1013
Inventor 王义翠张通李硕刘奕
Owner JINAN KAICHEN BIOTEC