Reagent for eliminating mycoplasma contamination in cell culture and use method thereof
A technology for removing cells and mycoplasma, which is applied in tissue culture, biochemical equipment and methods, and microbial measurement/inspection, and can solve the problems of reagents and methods that have not been studied for mycoplasma contamination.
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Embodiment 1
[0047]The reagent for removing mycoplasma contamination in cell culture is formed by mixing equal volumes of liquid A and liquid B, wherein:
[0048] Liquid A is formed by mixing tiamulin and 0.01MPBS, and the mass-volume concentration of tiamulin in the mixture is 1.5%;
[0049] Solution B is formed by mixing enrofloxacin and 0.01MPBS, and the mass-volume concentration of enrofloxacin in the mixed solution is 1%;
[0050] Mix equal volumes of liquid A and liquid B, and the mass ratio of tiamulin to enrofloxacin in the mixed liquid is 3:2.
[0051] A method for removing mycoplasma contamination with the above reagents, selects the cultivation of A549 cells (human lung adenocarcinoma cells) as the experimental object, and its concrete steps are as follows:
[0052] a. Mycoplasma detection
[0053] (1) The cell culture medium of A549 cells was prepared by adding 10% FBS (domestic Sijiqing fetal bovine, mycoplasma-free grade) into DMEM high-glucose medium (manufactured by Ameri...
Embodiment 2
[0069] The reagent for removing mycoplasma contamination in cell culture is formed by mixing equal volumes of liquid A and liquid B, wherein:
[0070] Liquid A is formed by mixing vornemulin and 0.01MPBS, and the mass-volume concentration of vornemulin in the mixed solution is 1.5%;
[0071] Liquid B is formed by mixing marbofloxacin and 0.01MPBS, and the mass-volume percentage concentration of marbofloxacin in the mixed solution is 1%;
[0072] Liquid A and liquid B are mixed in equal volumes, and the mass ratio of warnemulin to marbofloxacin in the mixed liquid is 3:2.
[0073] A kind of method that removes mycoplasma pollution with above reagent, selects the cultivation of SH-SY-5Y cell (human neuroblastoma) as experiment, and its concrete steps are as follows:
[0074] a. Mycoplasma detection
[0075] (1) For the cell culture medium of SH-SY-5Y cells, DMEM high-glucose medium (manufactured by GIBCO) and 10% FBS (domestic Sijiqing Tainiu, mycoplasma-free grade) were added...
Embodiment 3
[0092]The reagent for removing mycoplasma contamination in cell culture is formed by mixing equal volumes of A liquid, B liquid, and C liquid, wherein:
[0093] Liquid A is formed by mixing vornemulin and 0.01MPBS, and the mass-volume concentration of vornemulin in the mixed solution is 1.5%;
[0094] Liquid B is formed by mixing enrofloxacin and 0.01MPBS, and the mass-volume percentage concentration of enrofloxacin in the mixed liquid is 1%;
[0095] Liquid C is formed by mixing minocycline and 0.01MPBS, and the mass-volume percentage concentration of minocycline in the mixed solution is 0.5%;
[0096] Liquid A, liquid B, and liquid C are mixed in equal volumes, and the mass ratio of warnemulin, enrofloxacin, and minocycline in the mixed liquid is 3:2:1.
[0097] A method for removing mycoplasma contamination with the above reagents, selects the cultivation of HELA cells as an experiment, and its concrete steps are as follows:
[0098] a. Mycoplasma detection
[0099] (1) ...
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