Strain preservation method

Abstract

The invention relates to a strain preservation method. According to the method, strains are mixed with a cryoprotectant prepared from 10 percent of glycerin and 2 percent of a trehalose solution, then the strains are put into a cryopreservation tube box with 100 holes, marks are made on the box cover, and the cryopreservation tube box is placed in a refrigerator with the temperature of 80 DEG C below zero for cryopreservation. When the preserved strains are used, required strains in cryoprotectan tubes can be found quickly according to the marks on the cryopreservation box cover. After being taken out, the cryopreservation tubes are immediately put into a water bath with the temperature of 35-40 DEG C for quickly unfreezing through vibration till completely thawed, and the strains in the cryoprotectan tubes are immigrated to suitable media for cultivation. By adopting the strain preservation method provided by the invention for strain preservation, the preservation cost is reduced greatly, special equipment is not required to be purchased, the cryoprotectant used is cheap and easy to get, the preparation process of the cryoprotectan tubes is simple and short in time consumption, the survival rate and purity are high, and the stability is good within five generations.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, and in particular relates to a preservation method for protecting the stability and metabolic activity of strains under ultra-low temperature freezing conditions. Background technique [0002] Several preservation methods commonly used in industrial production include: regular transplantation method, liquid paraffin oil preservation method, sand tube preservation method, freeze vacuum drying method, liquid nitrogen ultra-low temperature freezing method, -80 ℃ low temperature freezing preservation and so on. Each of these preservation methods has its advantages and disadvantages. For example, in the regular transplantation method, the moisture in the culture medium of the slant is easy to evaporate and shrink, dry up, and collapse during the storage period, so that the concentration increases and the osmotic pressure increases, which will cause the degradation or even death of the ...

AI Technical Summary

Problems solved by technology

For example, in the regular transplantation method, the moisture in the culture medium of the slant is easy to evaporate and shrink, dry up, and collapse during the storage period, so that the concentration increases and the osmotic pressure increases, which will cause the degradation or even death of the strain; some favorable traits will be weakened after continuous passage; Easy to cause bacterial contamination

Liquid paraffin oil preservation method: most of the paraffin oil is chemically pure, if it contains impurities, especially toxic substances, it will affect the production performance of the strain when it is used for the preservation of the strain; during the preservation process, the paraffin oil will reduce due to evaporation, resulting in inclined If the culture is exposed, if it is not replenished in time, it will cause the strain to degenerate or even die; the slope for preservation must be placed upright, which occupies too much storage space and requires a lot of storage equipment

Sand tube preservation method: the soil as one of the storage carriers should be taken from barren soil, and the material has high requirements; sand and soil need to be washed, removed impurities, and iron filings; the proportion of sand in the sand tube is not fixed, and is affected by the quality of sand and bacteria. Affected by various characteristics, it often needs to be determined through experiments; low survival rate and high mutation rate

Freeze vacuum drying method: high requirements for equipment, long preparation time and high requirements for operators’ skills

Liquid nitrogen ultra-low temperature freezing method: It needs to be frozen slowly to -40°C first, and then quickly cooled to -150°C in a liquid nitrogen tank for storage; the operation is relatively cumbersome and requires high equipment requirements

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