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Method for preserving cordyceps militaris strain for long time

A long-term preservation, Cordyceps militaris technology, applied in the preservation of microorganisms and other directions, can solve the problems of needing to be placed upright, the paraffin preservation method is not suitable, and achieve the effects of ensuring dryness, inhibiting physiological activity, and inhibiting physiological activity.

Inactive Publication Date: 2015-03-25
东莞市生物技术研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is simple to make and does not require special equipment. The bacteria can be stored for about 1 year without frequent transplantation. The disadvantage is that it needs to be placed upright during storage.
[0003] However, for some strains with tenacious vitality, such as Cordyceps militaris strains, the existing paraffin preservation method is not applicable. Therefore, it is necessary to develop a long-term preservation method that can better inhibit the physiological activities of the strains

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A method for long-term preservation of Cordyceps militaris strain, comprising the steps:

[0024] A, bacterial culture: under aseptic conditions, the healthy strains of Cordyceps militaris are inoculated on the sterilized PDA slant medium, carry out dark culture, and after the mycelium is covered with 40% of the slope, carry out light culture;

[0025] B. Dehydration of the culture medium: put the test tube containing the culture into a drying dish, and then place the drying dish in the refrigerator. When the slanted medium is dehydrated to the remaining 20% ​​of the volume, take out the test tube;

[0026] C. Preservation of strains: slowly pour sterilized paraffin oil into the test tube on the ultra-clean workbench until the culture is submerged by 1cm, remove the bubbles on the surface of the culture with a small vortex shaker, insert the sterilized absorbent cotton, and use After sterilized paraffin seal, cover with sterilized pp film, and store in a refrigerator at...

Embodiment 2

[0032] A method for long-term preservation of Cordyceps militaris strain, comprising the steps:

[0033] A, bacterial culture: under aseptic conditions, the healthy strains of Cordyceps militaris are inoculated on the sterilized PDA slant medium, carry out dark culture, and after the mycelium is covered with 45% of the slope, carry out light culture;

[0034] B. Dehydration of the culture medium: put the test tube containing the culture into a drying dish, and then place the drying dish in the refrigerator. When the slanted medium is dehydrated to 22% of the volume remaining, take out the test tube;

[0035] C. Preservation of strains: slowly inject sterilized paraffin oil into the test tube on the ultra-clean workbench to submerge the culture for 1.5cm, remove the bubbles on the surface of the culture with a small vortex shaker, and insert the sterilized absorbent cotton. Seal with sterilized paraffin, cover with sterilized pp film, and store in a refrigerator at 0°C vertical...

Embodiment 3

[0041] A method for long-term preservation of Cordyceps militaris strain, comprising the steps:

[0042] A, bacterial culture: under aseptic conditions, the healthy strains of Cordyceps militaris are inoculated on the sterilized PDA slant medium, carry out dark culture, and after the mycelium is covered with 50% of the slope, carry out light culture;

[0043] B. Dehydration of the culture medium: put the test tube containing the culture into a drying dish, and then place the drying dish in the refrigerator. When the slanted medium is dehydrated to the remaining 25% of the volume, take out the test tube;

[0044] C. Preservation of strains: slowly inject sterilized paraffin oil into the test tube on the ultra-clean workbench to submerge the culture for 2 cm, use a small vortex shaker to remove the bubbles on the surface of the culture, insert the sterilized absorbent cotton, and use After sterilized paraffin seal, cover with sterilized pp film, and store in a refrigerator at 0°...

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PUM

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Abstract

The invention relates to the technical field of preservation of strains and in particular relates to a method for preserving a cordyceps militaris strain for a long time. The method comprises the following steps: A. culturing the strain, namely inoculating a healthy cordyceps militaris strain to a sterilized PDA inclined medium under aseptic condition, carrying out dark culture, and carrying out light culture after hyphae overgrows 40%-60% of the slope; B. dewatering the culture medium, namely putting a test tube filled with culture into a drying vessel, then putting the drying vessel in a refrigerator, and taking out the test tube after the inclined medium is dewatered until 20%-30% of volume is left; and C. preserving the strain, namely slowly injecting sterilized paraffin oil into the test tube till the paraffin oil immerses the culture by over 1cm, shaking and removing bubbles on the surface of the culture, plugging sterilized and degreased cotton, sealing by using sterilized paraffin, covering a sterilized pp film, vertically arranging the test tube in the refrigerator at 0 DEG C for preserving. By virtue of the method, the physiological activity of the strain can be well inhibited; the strain can be preserved for more than 3 years; the preserved strain also has good activity.

Description

technical field [0001] The invention relates to the technical field of strain preservation, in particular to a method for long-term preservation of Cordyceps militaris strains. Background technique [0002] The general steps of the existing paraffin preservation method are as follows: sterilize the liquid paraffin, place it in a 37° C. incubator, and evaporate the water vapor for use. Then, the purified bacteria to be stored are cultivated in the most suitable slant medium, so that a robust bacterial body is obtained, and the sterilized liquid paraffin is sucked with a sterile pipette and injected into the grown slant medium. The test tube is upright, labelled, and stored at 4°C. The method is simple to manufacture, does not require special equipment, and the strains can be stored for about one year without frequent transplanting. The disadvantage is that they need to be placed upright during storage. [0003] However, for some strains with tenacious vitality, such as Cor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04
CPCC12N1/04
Inventor 郑贵朝叶锦泉喻孟君
Owner 东莞市生物技术研究所
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