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Preservation method for bacteriophage

A preservation method and phage technology, which is applied in the field of biotechnology and microbial products, can solve the problems of restricting the application and development of phage, the short storage time of phage, and the poor preservation effect of phage, and achieve low storage environment requirements, good adaptability and convenient application Effect

Inactive Publication Date: 2016-09-21
SUZHOU IRIVET BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, in the above-mentioned process, the phages need to be preserved. In the preservation scheme of the prior art, the preservation effect of the phages is poor, the preservation time of the phages is short, the survival rate is low, the phages are easily contaminated, and must be preserved under low temperature conditions. Seriously restricts the development of phage applications

Method used

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  • Preservation method for bacteriophage
  • Preservation method for bacteriophage

Examples

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Embodiment 1

[0024] A preservation method of coliphage, completed under 4% humidity conditions, comprising the following steps:

[0025] 1) The E. coli phage solution was left standing at 2°C for 4 hours, flocculent precipitates appeared, and centrifuged at 8000r / min for 20min;

[0026] 2) Add phage buffer solution to the above-mentioned precipitate, dissolve the precipitate, and obtain phage liquid; the phage buffer solution includes the following components in terms of weight components: 8 parts of Tris hydrochloride, sodium chloride 0.05 parts, 1 part of potassium chloride, 5 parts of magnesium sulfate, 1 part of dipotassium hydrogen phosphate, 8 parts of sodium dihydrogen phosphate, 0.05 parts of gelatin and 100 parts of deionized water, and use dilute hydrochloric acid and / or sodium hydroxide to adjust the pH to 7;

[0027] 3) The phage preservation and protection agent equal to the volume of the phage liquid is mixed evenly to obtain the phage stock solution. The amino acids of the ...

Embodiment 2

[0031] A method for preserving Mycobacterium phage, which is completed under 6% humidity conditions, comprising the following steps:

[0032] 1) The Mycobacterium phage solution was left standing at 7°C for 8 hours, flocculent precipitates appeared, and centrifuged at 12000r / min for 40min;

[0033] 2) Add phage buffer solution to the above-mentioned precipitate, dissolve the precipitate, and obtain phage liquid; the phage buffer solution includes the following components in terms of weight components: 15 parts of Tris hydrochloride, sodium chloride 0.2 parts, potassium chloride 5 parts, magnesium sulfate 20 parts, dipotassium hydrogen phosphate 3 parts, sodium dihydrogen phosphate 12 parts, gelatin 0.1 part and purified water 100 parts, and use dilute hydrochloric acid and / or sodium hydroxide to adjust the pH to 7.5.

[0034] 3) A phage preservation and protection agent 2.5 times the volume of the phage liquid was mixed evenly to obtain a phage stock solution. The amino acids...

Embodiment 3

[0038] A method for preserving vibrio alginolyticus phage, which is completed under 5% humidity conditions, comprising the following steps:

[0039] 1) The Vibrio alginolyticus phage solution was left standing at 5°C for 6 hours, flocculent precipitates appeared, and centrifuged at 10,000 r / min for 30 minutes;

[0040] 2) Add phage buffer solution to the above-mentioned precipitate, dissolve the precipitate, and obtain phage liquid; the phage buffer solution includes the following components in terms of weight components: 12 parts of Tris hydrochloride, sodium chloride 0.1 parts, 3 parts of potassium chloride, 10 parts of magnesium sulfate, 2 parts of dipotassium hydrogen phosphate, 10 parts of sodium dihydrogen phosphate, 0.08 parts of gelatin and 100 parts of deionized water, and use dilute hydrochloric acid and / or sodium hydroxide to adjust the pH to 7.2;

[0041] 3) A phage preservation and protection agent with 2.2 times the volume of the phage liquid was mixed evenly to...

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Abstract

The invention provides a preservation method for bacteriophage. The preservation method comprises the steps that bacteriophage liquid and a bacteriophage preservation protecting agent are mixed to be uniform to obtain a bacteriophage stock solution, and then the bacteriophage stock solution is subpackaged into ampoules to be sealed and cryo-preserved at room temperature or the temperature of 2 DEG C to 7 DEG C. The preservation method for the bacteriophage is reasonable and convenient to apply, and the bacteriophage can be preserved at the room temperature or refrigerated, is low in requirement for the preservation environment and good in adaptability and can be preserved for a long term.

Description

technical field [0001] The invention belongs to the fields of biotechnology and microbial products, and in particular relates to a method for preserving bacteriophage. Background technique [0002] Phage is a general term for a class of viruses that can infect microorganisms such as bacteria, actinomycetes, and spirochetes. No toxicity to animals and plants. With the development of molecular biology technology, phage has been widely used in many fields such as disease diagnosis and water treatment. Phages have also become important molecular and genetic research tools due to their ability to insert genes into host DNA. Using phages, many ingenious experiments can be designed. [0003] However, in the above-mentioned process, the phages need to be preserved, the preservation effect of the phages in the preservation scheme of the prior art is poor, the preservation time of the phages is short, the survival rate is low, the phages are easily contaminated, and must be preserv...

Claims

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Application Information

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IPC IPC(8): C12N7/00
CPCC12N7/00C12N2795/00011
Inventor 许维素宋增福陈彪
Owner SUZHOU IRIVET BIOTECH
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