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Pretreatment kit and method for bacterial plasmid extraction

A bacterial plasmid and kit technology, which is applied in the field of pretreatment kits for bacterial plasmid extraction, can solve the problems of difficult plasmid extraction, high cross-linking degree, and high cost, and overcome the inability to effectively extract Gram-positive bacterial plasmids, Broad market application prospects, excellent extraction effect

Active Publication Date: 2018-11-23
CHENGDU MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Because of the peptidoglycan in the cell wall of Gram-positive bacteria, the degree of cross-linking is high, and it is difficult to cleavage, which makes the plasmid extraction of Gram-positive bacteria very difficult, especially one or several natural plasmids in many Lactobacillus plantarum strains, extraction is more difficult
It has been reported in the existing literature that glass beads combined with short vortexing time can help overcome the difficulty of lysis, but the operation is complicated and costly

Method used

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  • Pretreatment kit and method for bacterial plasmid extraction
  • Pretreatment kit and method for bacterial plasmid extraction
  • Pretreatment kit and method for bacterial plasmid extraction

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Embodiment 1

[0029] Embodiment 1, the pretreatment kit and method of bacterial plasmid extraction of the present invention

[0030] 1. Kit composition

[0031] A component: 100mg / ml lysozyme

[0032] B component: 5% (refers to 5g / 100ml) glucose solution

[0033] 2. How to use

[0034] Take the bacteria, add 100 mg / ml lysozyme, and incubate at 37°C for 10 minutes. After lysozyme treatment, centrifuge the suspension at 11,000×g for 1 minute at room temperature; Gently suspend in 5% glucose solution, centrifuge at 11,000×g for 1 minute, and discard the supernatant.

Embodiment 2

[0035] Embodiment 2, the pretreatment kit and method of bacterial plasmid extraction of the present invention

[0036] 1. Kit composition

[0037] Component A: 120mg / ml lysozyme

[0038] Component B: 4% glucose solution

[0039] 2. How to use

[0040] Take the bacteria, add 150 mg / ml lysozyme, and incubate at 37°C for 10 minutes. After lysozyme treatment, centrifuge the suspension at 11,000×g for 1 minute at room temperature; Gently suspend in 5% glucose solution, centrifuge at 11,000×g for 1 minute, and discard the supernatant.

Embodiment 3

[0041] Embodiment 3, the pretreatment kit and method of bacterial plasmid extraction of the present invention

[0042] 1. Kit composition

[0043] A component: 160mg / ml lysozyme

[0044] Component B: 5% glucose solution

[0045] 2. How to use

[0046] Take the bacteria, add 200mg / ml lysozyme, and incubate at 37°C for 10 minutes. After lysozyme treatment, centrifuge the suspension at 11,000×g for 1 minute at room temperature; Gently suspend in 5% glucose solution, centrifuge at 11,000×g for 1 minute, and discard the supernatant.

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Abstract

The invention relates to a pretreatment kit for bacterial plasmid extraction. The pretreatment kit comprises a component A, namely 100-200mg / ml lysozyme, and a component B, namely a washing solution,wherein preferably, the component A is 100mg / ml lysozyme. The invention also discloses a pretreatment method for bacterial plasmid extraction, and a bacterial plasmid extraction kit and method. The pretreatment kit and method for the bacterial plasmid extraction can effectively improve the bacterial plasmid extraction efficiency, particularly have greatly excellent lactobacillus plantarum extraction effect, can solve the problem of being incapable of efficiently extracting gram-positive bacterium plasmid of existing methods, and thus have broad market application prospects.

Description

technical field [0001] The invention relates to a pretreatment kit and method for bacterial plasmid extraction. Background technique [0002] Plasmids are nucleic acid molecules capable of self-replicating in living cells. Plasmid-mediated horizontal gene transfer is considered a major driver of bacterial diversity and adaptation. With the deepening of research, plasmids have been widely used in genetic research tools. The extraction of plasmid DNA is a key technique in molecular biology and an essential link in molecular techniques such as cloning, transfection and gene therapy. New plasmids have been found to play a very important role in bacteria. [0003] Because of the peptidoglycan in the cell wall of Gram-positive bacteria, the degree of cross-linking is high, and it is difficult to cleavage, which makes the plasmid extraction of Gram-positive bacteria very difficult, especially one or several natural plasmids in many Lactobacillus plantarum strains, Extraction is...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q2527/125
Inventor 潘渠姚芳许晓羽杜昕曹康
Owner CHENGDU MEDICAL COLLEGE
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