Liquid nitrogen preservation method, also known as cryopreservation method. It mainly utilizes the principle that microorganisms tend to stop at a low temperature below -130 ° C, and effectively preserves the strain in liquid nitrogen (at an ultra-low temperature of -196 ° C). The method is simple and efficient in operation, and the preservation period can generally reach more than 15 years. It is one of the currently recognized long-term preservation techniques for strains and is suitable for the preservation of various microbial strains. However, the use of liquid nitrogen preservation requires the purchase of ultra-low temperature liquid nitrogen equipment, and the liquid nitrogen consumption is large and the cost is high. Liquid nitrogen cryopreservation protection agent, generally choose glycerin, dimethyl sulfoxide, dextrin, serum protein, polyethylene nitrogen ring, Tween 80, etc., the most commonly used is glycerol (10% ~ 20%), different microorganisms Different protective agents should be chosen, and the concentration of the protective agent should be controlled so as not to cause microbial death.
1. Simple preservation method, agar slant spore culture, mycelial suspension and spore culture made from cereal materials such as bran, rice and millet are stored in a refrigerator at 4 ° C for less than 1 to 2 months. If the spore bottle prepared from the grain raw material is vacuumed and immersed in the cotton plug to isolate the outside air and water vapor, the holding time can be 3 to 4 months.
2. Liquid nitrogen ultra-low temperature preservation method, suspending the cells in the stable growth period in 10% glycerin or other low freezing point liquid, sealing in the ampoule tube, and then controlling the cooling rate, so that the temperature of the ampoule tube is gradually lowered to -35 ° C, then Store in a liquid nitrogen tank at -150 to -196 °C. Most microorganisms such as viruses, bacteriophages, various bacteria, actinomycetes, yeasts and protozoa, especially those that have difficulty in freeze-drying, can be preserved for a long time.
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