Subculture of tissue culture

[Method steps]
1 Take materials. In the clean workbench in the sterile room, take a bottle of culture at a time, burn about 20 mm from the mouth of the culture bottle, and use the tweezers and scalpel after burning to remove the explants or calluses. The tissue is placed in a sterile petri dish. Approximately 4-6 explants or callus are placed in each petri dish. Divide each callus into two or three small pieces not less than 5 mm square. The cells below the explant toward the center are often necrotic, which is not suitable for subculture. These necrotic parts should be separated from the normal parts and discarded. After each operation, remove the used absorbent paper and replace the lid of the petri dish.
2 Subsequent transfer. Transfer the developing explants or callus to fresh medium, and the inoculation method is the same as before. After the transfer, write the culture, medium, and date on the culture flask. Long-term cultivation of carrot tissue will produce large pieces of callus. The experimental procedure used for the first subculture of the developing callus is the same as the subculture procedure of transferring the established callus line every four weeks in the future.
3 Experiment records. Copy the experiment records in a notebook, and indicate the date of the experiment, the duration, the number of cultures, the number of contamination, and the different treatments done. During four weeks, observe the culture with the naked eye every other week, record the changes in the morphology of the culture, the growth state of the culture, the change in fresh weight, etc., and take pictures if necessary.

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