Liquid strain is a strain cultured with liquid culture medium. It has the advantages of short strain production cycle, consistent bacterial age, convenient inoculation, suitable for factory production, and so on. It has been welcomed by the majority of growers. Cell shaker （https://www.luoron.com/plastic-erlenmeyer-flask-with-vent-cap-2-product/） is a kind of consumable material needed in the culture of liquid strains. The following points need to be paid attention to in the culture:

1.Preparation of culture medium according to the formula of various components, into the container, dissolve with water, divided into triangular bottles, generally 250 ~ 300mL cell shake bottle 50mL culture medium, 500mL triangle bottle  （https://www.luoron.com/plastic-erlenmeyer-flask-with-vent-cap-2-product/）100mL culture medium, plug cotton plug, with newspaper wrapped; Can also be made of 8 layers of gauze 8cm×8cm bottle stopper sealing, outside with brown paper sealing.
2. Autoclave sterilization method is generally adopted for sterilization, which requires 121℃ and 30min. Take it out and cool it to about 30℃. Put it in the sterile room or inoculation box for use.
3. According to aseptic operation requirements, each bottle can be connected with a 2 ~ 3cm2 slant strain, and each slant strain can be connected with 4 ~ 5 bottles. It is better to bring some medium with the added strains, so that they can float on the surface of the medium. After inoculation, cover the mouth of the bottle with the gauze originally plugged, and tie it firmly with string.
4. The inoculated strain bottle can be placed in a shaking bed for culture, or can be placed in a constant temperature of 24 ~ 26℃ for 48h, and then oscillated culture can be carried out after the aerogenic mycelium is extended into the culture medium. The oscillating frequency of reciprocating shaker is 80 ~ 120r/min, and that of rotary shaker is 150 ~ 220r/min. 3 ~ 4d can be cultured in the shaker. At the end of culture, due to different strains, the culture medium appeared different colors, such as oyster mushroom, enoki mushroom culture medium was light yellow; Mushroom, ericius erinaceus culture medium was reddish brown, and mushroom fragrance; The fungus culture medium is brown, thick and sweet. The culture medium can be used for production or further expansion culture only when the dry weight of mycelium reaches 10g/L and the diameter of mycelium pellets is 1 ~ 2mm.
 
5. Secondary liquid strain production secondary liquid strain culture medium production of the same level of liquid strain, the culture container should be larger, the capacity is not more than 3500mL. After sterilization and cooling, the fermented primary liquid strains were inserted into 5L cell shaker （https://www.luoron.com/3l5l-high-efficiency-erlenmeyer-flask-product/）at a ratio of 5%-10% and cultured on a shaker at a slow speed. After a certain time of oscillating culture, the liquid strains with uniform distribution of microspheres and clear fermentation liquid can be obtained.
In summary, when using cell shaker to culture liquid strains, operate according to the above methods, and pay attention to control the temperature, shaking frequency and other factors in the process to ensure the culture effect.
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