Q1: In step 3, why after 6-12 hours when ~80% are cleared, should the medium be changed once?
It can be tried without changing the medium. It does no harm to the cells. It is recommended to change the liquid according to our quality control operation, just to remove the dead black glue bugs in time. So as to make the subsequent cultivation cleaner
Q2: In step 5, the instructions suggest that after removing the black worms, continue to add reagent B to the medium during subsequent culture. For how long should this be done?
The addition can be stopped at any time, but since the black glue bug is prone to resurface, it is recommended to add reagent B for more than 10 generations or continue until the end of the experiment. All these are fine.
Q3: Can it be used for suspension cells?
Regrettably, we haven't conducted any tests in this regard so far. Theoretically, it can be attempted. In terms of method, for suspension cells, since they do not adhere, all the instructions in the manual regarding before and after adhesion are meaningless. It is recommended to directly add the black gum bug removal reagent at the concentration specified in the instructions to the culture medium. Reagent A and Reagent B should be added simultaneously. Similarly, the addition of Reagent A should not exceed three generations, while reagent B can be added continuously.
Q4: My cells need to be passaged very infrequently. The instructions say to add it only during passaging. Now they are contaminated. Can I add it without passaging?
It can be added without subculture and has no effect on the cells, but it may not have a good killing effect on black glue worms. In cases like this, it is recommended to subculture at a ratio of 1:1 or 1:0.5. That is to say, the cells need to be digested and reintroduced into a new well of the same area or a smaller well (for example, one well in a six-well plate is digested and then one well in a 12-well plate), allowing the cells to reattach. Adding chemicals for extermination during this process will yield better results. In addition, if the cells are subcultured very frequently, it is recommended that the addition time of Reagent A should not be too long. For example, remove it after 24 to 48 hours of treatment and replace it with a new medium. Then, continuously add Reagent B to the new medium.
Q5: When adding the black worm remover, do I need to temporarily stop using antibiotics?
The bispecific antibody has no cross-reaction with our reagent and does not need to be removed.
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