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Liquid nitrogen cryopreservation is the most effective and reliable method for long-term preservation of bacteria

Time: 2019-06-05 10:55 Source: unknown Author: Bander liquid nitrogen tank Click: second
How to use liquid nitrogen preservation? Self pressurized liquid nitrogen tank

The liquid cryopreservation method is to put the bacteria in the ampoule containing cryoprotectant and put the ampoule in liquid nitrogen (- 196 ℃). As the mycelium was at - 196 ℃, its metabolism was reduced to the state of complete stop.

Therefore, there is no need to transplant them regularly.

Liquid nitrogen cryopreservation is the most effective and reliable method for long-term preservation of bacteria.

The specific operation is as follows:

(1) Strain preparation: the strain was cultured on PDA medium plate at 22-24 ℃ for 10-15 days. After the mycelium was fully grown, agar block was cut with a puncher (2-4 mm in diameter), or the Mycelium Culture was cut into 2 mm x 4 mm rectangular pieces with surgical blade.

(2) The glass of the bottle fixing tube should be able to withstand the sudden change of temperature without breaking. It is easy to seal the tube mouth with flame and open easily when the culture is restored. Borosilicate glass is generally used. The size of the tube depends on the needs. It is usually 75 mm x 10 mm or an ampoule that can hold 1.2 ml liquid.
After the ampoule tube is selected, the number of the edible fungus species to be preserved is written on the tube wall with ink, and then sterilized by dry heat at 160 ℃, so that the mark is firmly fixed on the tube wall.    

(3) Protective agent: add 0.8 ml of autoclaved 10% glycerin aqueous solution or 10% dimethylene aqueous solution into each tube, which can be used as cryoprotectant.
The agar block with mycelium was transferred into an ampoule with protective agent by sterile tweezers. Melt the upper part of the ampoule tube with flame and immerse it in water to check whether there is air leakage.

(4) In liquid nitrogen storage, put the finished strain tube into a small plastic bottle with holes (place a piece of iron at the bottom of the plastic bottle, so that the plastic bottle and the ampoule can sink together liquid nitrogen tank Put the cap on the bottle, connect a string on the cap and mark it, so as to put forward the required strain. Hang the plastic bottle with an ampoule in the liquid nitrogen tank After about 30 minutes, immerse the ampoule in liquid nitrogen for long-term preservation.

When inoculating the bacteria in the culture flask at the temperature of 22-40 ℃, the bacteria should be put into the culture flask at the temperature of 35-40 ℃, and then put the bacteria into the culture flask at suitable temperature of 35-40 ℃. Self pressurized liquid nitrogen tank

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