Mycaway plus color one step mycoplasma detection kit

Manufactured by Yeasen

Sourced in China

MycAwayTM Plus-Color One-Step Mycoplasma Detection Kit is a laboratory tool used for the detection of mycoplasma contamination in cell cultures. The kit provides a one-step procedure to identify the presence of mycoplasma DNA.

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Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Primary normal human astrocytes nhas, by ScienCell (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Sangon (1 mentions) Fetal bovine serum (fbs), by Transgene (1 mentions) U251mg, by American Type Culture Collection (1 mentions) Hek293t, by American Type Culture Collection (1 mentions) U87mg, by American Type Culture Collection (1 mentions)

6 protocols using mycaway plus color one step mycoplasma detection kit

Standardized Cell Culture Protocols

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Detailed information on HOK, UM1, UM2, HSC-3, CAL27, HN12, HN31, HN30, H413, and HEK293T cells is shown in Appendix Table S3. HOK cells were cultured in a defined keratinocyte SFM medium (10744019, Thermo Fisher Scientific). UM1, UM2, HSC-3, CAL27, HN12, HN31, H413, and HEK293T cells were appropriately cultured in DMEM (SH30243.01, HyClone) supplemented with 10% FBS and 1% penicillin-streptavidin solution. All cells were regularly tested for mycoplasma with a MycAwayTM Plus-Color One-Step Mycoplasma Detection Kit (40612ES25, Yeasen). Cells were kept in a humidified incubator at 37 °C with 5% CO₂.

Wei Z., Wang Y., Peng J., Li H., Gu J., Ji N., Li T., Zhou X., Zeng X., Li J, & Chen Q. (2022). CircRFWD3 promotes HNSCC metastasis by modulating miR-27a/b/PPARγ signaling. Cell Death Discovery, 8, 285.

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Cell Culture Protocols for Cancer Research

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HEK293T, HEB, U-251MG (U251), U-118MG (U118), U87MG (U87) cells were purchased from the Cell Bank of the Chinese Academy of Sciences (Chinese Academy of Sciences, Shanghai, China). Cells were cultured in DMEM/High Glucose (HyClone, USA) supplemented with 10% FBS and 1% penicillin/streptomycin at 5% CO₂ at 37 °C. All cell lines were routinely screened for mycoplasma contamination by MycAwayTM Plus-Color One-Step Mycoplasma Detection Kit (Yeasen, Shanghai, China).

Dong N., Qi W., Wu L., Li J., Zhang X., Wu H., Zhang W., Jiang J., Zhang S., Fu W., Liu Q., Qi G., Wang L., Lu Y., Luo J., Kong Y., Liu Y., Zhao R.C, & Wang J. (2024). LINC00606 promotes glioblastoma progression through sponge miR-486-3p and interaction with ATP11B. Journal of Experimental & Clinical Cancer Research : CR, 43, 139.

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Glioblastoma Cell Line Cultivation

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Human U87, U251, A172, and U373 glioblastoma cell lines were purchased from the ATCC (Manassas, VA, USA). Primary normal human astrocytes (NHAs) were from Sciencell Research Laboratories (Carlsbad, CA, USA). NHAs were cultured in Dulbecco’s modified Eagle’s medium (DMEM), while all cultured glioma cell lines were cultured in DMEM-F12. Both mediums were supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY, USA) and incubated at 37°C and 5% CO₂. No mycoplasma contamination was detected in all cell lines by the MycAwayTM Plus-Color One-Step Mycoplasma Detection Kit (Yeasen, Shanghai, China).

Wu Z., Zheng M., Zhang Y., Xie M., Tian S., Ding T., Li L, & Guan Q. (2020). Hsa_circ_0043278 functions as competitive endogenous RNA to enhance glioblastoma multiforme progression by sponging miR-638. Aging (Albany NY), 12(21), 21114-21128.

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Cell Culture and Mycoplasma Testing

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HEK293T and CRC cells were purchased from the Type Culture Collection of the Chinese Academy of Sciences. DLD-1 cells were cultured in RPMI-1640 (Gibco) supplemented with 10% fetal bovine serum (FBS; Anlite) and 1% penicillin-streptomycin (Sangon Biotech). Except for DLD-1, all the other cell lines were cultured in DMEM (Gibco). Cells were incubated at 37°C in a humidified 5% CO 2 atmosphere. All cells were frozen and stored in liquid nitrogen before experimentation. Cells were brought into culture for no more than 4 wk. Mycoplasma testing was performed approximately every 3 mo on all cells in use at the time using MycAway Plus-Color One-Step Mycoplasma Detection Kit from Yeasen Biotechnology.

Xia J., Ma N., Shi Q., Liu Q.C., Zhang W., Cao H.J., Wang Y.K., Zheng Q.W., Ni Q.Z., Xu S., Zhu B., Qiu X.S., Ding K., Huang J.Y., Liang X., Chen Y., Xiang Y.J., Zhang X.R., Qiu L., Chen W., Xie D., Wang X., Long L, & Li J.J. (2024). XAF1 promotes colorectal cancer metastasis via VCP-RNF114-JUP axis. The Journal of cell biology, 223(2).

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Cell Line Cultivation and Mycoplasma Screening

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U87 MG, U251 MG, T98G and HEK293T cell lines were purchased from ATCC and cultured in Dulbecco’s Modified Eagle’s Medium (Gibco, Cat. no. C11995500BT) supplemented with 10% fetal bovine serum (TransGen Biotech, Cat. no. FS301-02) and 1% penicillin and streptomycin (Aladdin, Cat. no. P301861-100 ml) at 37 ℃and 5% CO₂. All cell lines were mycoplasma negative detected by MycAway™ Plus-Color One-Step Mycoplasma Detection Kit (Yeasen, Cat. no. 40612ES25).

Li X., Sun S., Zhang W., Liang Z., Fang Y., Sun T., Wan Y., Ma X., Zhang S., Xu Y, & Tian R. (2024). Identification of genetic modifiers enhancing B7-H3-targeting CAR T cell therapy against glioblastoma through large-scale CRISPRi screening. Journal of Experimental & Clinical Cancer Research : CR, 43, 95.

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Hypoxia-Inducible Factor-1 Alpha Expression

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The CDs of Hif-1α1 and Hif-1α2 were cloned into pcDNA3.1 expression vector with Flag-tag on the C-terminal ends of the target genes. The HEK293T cells (RRID: CVCL_0063) were seeded in 2 mL of DMEM (Thermo Fisher) in a six-well plate 1 day before transfection. The Hif-1α1 or Hif-1α2 expression vectors (1 µg/well) were transfected to the HEK293T cells by using Lipofectamine 3000 (Thermo Fisher, L3000015). The transfected cells were cultured for additional 36 hr at 37°C for protein expression tests. For hypoxia treatment, the transfected cells were cultured for 30 hr at normoxia (5% CO₂ and 95% air) and additional 6 hr at hypoxia (1% O₂, 5% CO₂, and 94% N₂) before the tests. The primers are listed in Supplementary file 1. The mycoplasma contamination of the HEK293T cell line was tested using the MycAway Plus-Color One-Step Mycoplasma Detection Kit (Yeasen Biotechnology, 40612ES60) once a month.

Ding D., Zhang J., Du B., Wang X., Hou L., Guo S., Chen B, & Kang L. (2022). Non-canonical function of an Hif-1α splice variant contributes to the sustained flight of locusts. eLife, 11, e74554.

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