PCR Mycoplasma Detection Kit

G238100 Reactions


DescriptionMycoplasma detection is often difficult due to its lack of visable appearance, therefore it can be afflicting your valuable cell and affecting your results without your knowledge. Cells contaminated by mycoplasma species can have changes in proliferation, metabolism, gene synthesis and processing, and even adhesion properties. The solution for quick and easy mycoplasma detection is ABM's PCR Mycoplasma Detection Kit. The PCR Mycoplasma Detection Kit allows for fast and reliable identification of mycoplasma contamination in cell cultures. Mycoplasma DNA in the cell culture supernatant is amplified via PCR and visualized using gel electrophoresis. In addition to the short detection process (less than 2 hours), the easy handling and high sensitivity makes this PCR Mycoplasma Detection Kit a convenient tool for routine examination of cell cultures and media.
Unit quantity100 Reactions

Do you need any kind of initial training to use your kit?
This kit is PCR based and very easy to use. There is no initial training for that. But we will be more than happy to answer any questions related to the use of this kit.
Do you provide or sell a positive control so that we can be sure the assay worked?
Yes, we do have a positive control included in the kit.
What size plate, dish or flask should I use for the culturing of my cells?
We recommend culturing your cells in a 6-well plate or 10cm dish.
What is the basis of the positive control in the mycoplasma kit?
It is DNA fragments containing proprietary sequences diluted in medium.
I noticed the product arrived with precipitation. Is that normal?
That is normal as the contents are concentrated. You may warm the product in a 37 degrees Celsius water bath until the sample has completely dissolved.
Can this kit be used for DNA samples extracted from cell pellets?
Yes, this kit will also work for extracted DNA samples
I kept the cells in the presence of gentamycin, Should I use antibiotics-free media to detect mycoplasma in the media?
It is not necessary to remove routine antibiotics (e.g. penicillin, streptomycin, and gentamicin) from the media prior to PCR detection, as they will not interfere with the assay.
What is the detection limit of your kit?
Our kit is able to detect as low as 10 copies of Mycoplasma /sample
What is the % of agarose gel that would work best for the G238 kit?
You may use standard 1% agarose gel and this will be the most cost-effective option. The product size is small (less than 500 bp) so 1.5% to 2% gel can also be used.
Our cells are grown in suspension and would not achieve 80% confluence, is there a target cell density (cells/ml) we should achieve to ensure we will detect the mycoplasma?
Mycoplasma testing is more dependent on how recently the cells have been subcultured. Seeding density does not play a major role when using this kit. We recommend cells be around at least 3-5 days old in culture without changing any fresh media so as to detect secreted mycoplasma in culture supernatant (i.e. the cell culture medium should not be refreshed/changed for 48-72 hours prior to collection).
Can I use cryogenically frozen supernatant?
Using samples stored at -80C should be fine for this kit. If you thawing out the cells and performing mycoplasma testing right away, usually when thawing the cells the supernatant from the vial is diluted when adding to the cell culture vessel. As long as the DMSO is diluted and the DMSO content is less than 0.5% final in the PCR reaction it should not cause a problem. Too much DMSO would alter the Tm of the PCR buffer and thus affect amplification.
What suggestions can you provide for the gel electrophoresis step?
We normally use 0.5X TAE buffer to cast 1% gel and also use the same buffer to run the electrophoresis in-house at 100V for about 20 minutes.
Is the positive control a mixture of more than one strain of mycoplasma DNA or inactivated mycoplasma that cannot divide?
The Mycoplasma positive control is a cloned vector containing a section of Mycoplasma gDNA sequence.
Since I have such a large number of cell lines to test, can I store the cell culture supernatant prior to use in the PCR, or does it have to be freshly collected?
In general, the more fresh the sample is when running the PCR, the better; in keeping with this, it is normally advised to wait until all cell lines are ready to go prior to collecting the supernatant for each. However, if necessary, you can certainly store collected supernatant at +4C storage for short-term (1-2 days) storage; note that we highly recommend avoiding repeated freeze thaw cycles.
I noticed that there are clear crystals in the product solution. Is it still functional?
The qPCR mastermix should still be functional. The crystals are from our proprietary buffering salt, which will be dissolved eventually when you warm the solution up to room temperature or even 37C. All our qPCR mastermixes are very stable and can withstand this kind of warming cycle. Once the salt is fully dissolved, the vial can be kept at 4C for 2-3 months.
How many days after thawing my cells can I perform this mycoplasma test?
We recommend the cells should remain in culture for at least 48-72 hours prior to screening for the presence of mycoplasmas, and that the media sample collection only be done once the cells have reached at least 80% confluence.
What size dish or flasks do you recommend the cells should be cultured in?
We encourage customers to culture their cells in either a 6-well plate or 10 cm plate.

  • Koh, JH et al. "Effect of water-soluble fraction of cherry tomatoes on the adhesion of probiotics and Salmonella to intestinal epithelial cells" J Sci Food Agric : (2013). PubMed: 23749725. Application: Cell Culture.
  • Jagadish, N et al. "A-kinase anchor protein 4 (AKAP4) a promising therapeutic target of colorectal cancer" J Exp Clin Cancer Res 34(1):142 (2015). DOI: 10.1186/s13046-015-0258-y. PubMed: 26590805. Application: Cell Culture.
  • Saha, A et al. "Effect of Metformin, Rapamycin, and Their Combination on Growth and Progression of Prostate Tumors in HiMyc Mice." Cancer Prev Res (Phila) 8(7):597-606 (2015). DOI: 10.1158/1940-6207.CAPR-15-0014. PubMed: 25908508. Application: Cell Culture.
  • Black, L. A et al. "In vitro activity of chloramphenicol, florfenicol and enrofloxacin against Chlamydia pecorum isolated from koalas (Phascolarctos cinereus)" Australian veterinary journal 11:420-423 (2015). DOI: DOI: 10.1111/avj.12364. Application: Mycoplasma detection.
  • Jagadish, N et al. "Heat shock protein 70–2 (HSP70-2) is a novel therapeutic target for colorectal cancer and is associated with tumor growth" BMC Cancer 1:561 (2016). DOI: 10.1186/s12885-016-2592-7. Application: Mycoplasma detection.
  • Blouin, M. J., Parisotto, M., Papavasiliou, V., Lavoie, C., Larsson, O., Ohh, M., ... & Ferbeyre, G. "Translational and HIF-1a-Dependent Metabolic Reprogramming Underpin Metabolic Plasticity and Responses to Kinase Inhibitors and Biguanides" Cell Metabolism 28:1-16 (2018).
  • Bufalieri, F., Infante, P., Bernardi, F., Caimano, M., Romania, P., Moretti, M., … Di Marcotullio, L. "ERAP1 promotes Hedgehog-dependent tumorigenesis by controlling USP47-mediated degradation of βTrCP" Nature Communications 10(1): (2019). DOI: 10.1038/s41467-019-11093-0.
  • Carter, B. Z., Mak, P. Y., Wang, X., Tao, W., Ruvolo, V., Mak, D., … Andreeff, M. "An ARC-Regulated IL1β/Cox-2/PGE2/β-Catenin/ARC Circuit Controls Leukemia–Microenvironment Interactions and Confers Drug Resistance in AML" Cancer Research 79(6):1165–1177 (2019). DOI: 10.1158/0008-5472.can-18-0921.
  • Chakravorty, S., Yan, B., Wang, C., Wang, L., Quaid, J. T., Lin, C. F., ... & Chopra, G. "Integrated Pan-Cancer Map of EBV-Associated Neoplasms Reveals Functional Host–Virus Interactions"  Cancer research. : (2019).
  • Guarino, A.M., Mauro, G.D., Ruggiero, G. "doi:10" 1038/s41598-019-45468-6 : (2019). DOI: 10.1038/s41598-019-45468-6.
  • Hsu, H. P., Wang, C. Y., Hsieh, P. Y., Fang, J. H., & Chen, Y. L. "Knockdown of serine/threonine-protein kinase 24 promotes tumorigenesis and myeloid-derived suppressor cell expansion in an orthotopic immunocompetent gastric cancer animal model" Journal of Cancer 11(1):213-228 (2020).
  • Hsueh, Y. S., Chang, H. H., Shan, Y. S., Sun, H. S., Fletcher, J. A., Li, C. F., & Chen, L. T. "Nuclear KIT induces a NFKBIB-RELA-KIT autoregulatory loop in imatinib-resistant gastrointestinal stromal tumors" Oncogene 38(38):6550-6565 (2019).
  • Ide, H., Inoue, S., Mizushima, T., Kashiwagi, E., Zheng, Y., & Miyamoto, H. "Role of glucocorticoid signaling in urothelial tumorigenesis: Inhibition by prednisone presumably through inducing glucocorticoid receptor transrepression" Molecular carcinogenesis. : (2019).
  • Inoue, S., Ide, H., Mizushima, T., Jiang, G., Netto, G. J., Gotoh, M., & Miyamoto, H. "Nuclear Factor-κB Promotes Urothelial Tumorigenesis and Cancer Progression via Cooperation with Androgen Receptor Signaling" Molecular Cancer Therapeutics 17(6):1303–1314 (2018). DOI: 10.1158/1535-7163.mct-17-0786.
  • Inoue, S., Mizushima, T., Ide, H., Jiang, G., Goto, T., Nagata, Y., … Miyamoto, H. "ATF2 promotes urothelial cancer outgrowth via cooperation with androgen receptor signaling" Endocrine Connections 1397–1408: (2018). DOI: 10.1530/ec-18-0364.
  • Jones, P. S., Yekula, A., Lansbury, E., Small, J. L., Ayinon, C., Mordecai, S., ... & Ghiran, I. "Characterization of plasma-derived protoporphyrin-IX-positive extracellular vesicles following 5-ALA use in patients with malignant glioma" EBioMedicine. : (2019).
  • Kollara, A., Shathasivam, P., Park, S., Ringuette, M. J., & Brown, T. J. "Increased Androgen Receptor Levels and Signaling in Ovarian Cancer Cells by VEPH1 Associated with Suppression of SMAD3 and AKT Activation" The Journal of Steroid Biochemistry and Molecular Biology 105498: (2019).
  • Lee, R. S., Zhang, L., Berger, A., Lawrence, M. G., Song, J., Niranjan, B., ... & Risbridger, G. P. "Characterization of the ERG-regulated Kinome in Prostate Cancer Identifies TNIK as a Potential Therapeutic Target" Neoplasia 21(4):389-400 (2019).
  • Linley, A. J., Griffin, R., Cicconi, S., D'Avola, A., MacEwan, D. J., Pettitt, A. R., ... & Slupsky, J. "Kinobead profiling reveals reprogramming of B-cell receptor signaling in response to therapy within primary CLL cells" bioRxiv 841312: (2019).
  • Lospinoso Severini, L., Quaglio, D., Basili, I., Ghirga, F., Bufalieri, F., Caimano, M., ... & Maroder, M. "A Smo/Gli Multitarget Hedgehog Pathway Inhibitor Impairs Tumor Growth" Cancers 11(10):1518 (2019).
  • Mehrpour, M., Rodriguez, R., Hamai, A., & Trang, M. "U" S. Patent Application No. 16/082 970: (2019).
  • Nii, T., Prabhu, V.V, Ruvolo, V., Madhukar, N., Zhao, R., Mu, H., Heese, L., Nishida, Y., Kojima, K., Garnett, M.J., McDermott, U., Benes, C.H., Charter, N., Deacon, S., Elemento, O., Allen, J.E., Oster, W., Stogniew, M., Ishizawa, J., Andreeff, M. "Imipridone ONC212 activates orphan G protein-coupled receptor GPR132 and integrated stress response in acute myeloid leukemia" Leukemia. : (2019). DOI: 10.1038/s41375-019-0491-z.
  • Ong, F., Dreesen, O., & Dröge, P. . "The RNA interactome of human telomerase RNA reveals a coding-independent role for 2 a histone mRNA in telomere homeostasis 3" : ().
  • Rosenberger, L., Ezquer, M., Lillo-Vera, F., Pedraza, P. L., Ortúzar, M. I., González, P. L., … Alcayaga-Miranda, F. "Stem cell exosomes inhibit angiogenesis and tumor growth of oral squamous cell carcinoma" Scientific Reports 9(1): (2019). DOI: 10.1038/s41598-018-36855-6.
  • Ross, J., Bressler, K., & Thakor, N. "Eukaryotic Initiation Factor 5B (eIF5B) Cooperates with eIF1A and eIF5 to Facilitate uORF2-Mediated Repression of ATF4 Translation" International Journal of Molecular Sciences 19(12):4032 (2018). DOI: 10.3390/ijms19124032.
  • Salvati, A., Gigantino, V., Nassa, G., Giurato, G., Alexandrova, E., Rizzo, F., … Weisz, A. "The Histone Methyltransferase DOT1L Is a Functional Component of Estrogen Receptor Alpha Signaling in Ovarian Cancer Cells" Cancers 11(11):1720 (2019). DOI: 10.3390/cancers11111720.
  • Severini, L. L., Quaglio, D., Basili, I., Ghirga, F., Bufalieri, F., Caimano, M., ... & Maroder, M. "A Smo/Gli Multitarget Hedgehog Pathway Inhibitor Impairs Tumor Growth" Cancers 11(10): (2019).
  • Shishodia, G., Koul, S., & Koul, H. K. "Protocadherin 7 is overexpressed in castration resistant prostate cancer and promotes aberrant MEK and AKT signaling" The Prostate 79(15):1739–1751 (2019). DOI: 10.1002/pros.23898.
  • Singh, S., Kumar, M., Kumar, S., Sen, S., Upadhyay, P., Bhattacharjee, S., ... & Kundu, T. K. "The cancer-associated, gain-of-function TP53 variant P152Lp53 activates multiple signaling pathways implicated in tumorigenesis" Journal of Biological Chemistry 294(38):14081-14095 (2019).
  • Spiombi, E., Angrisani, A., Fonte, S., De Feudis, G., Fabretti, F., Cucchi, D., ... & Di Magno, L. "KCTD15 inhibits the Hedgehog pathway in Medulloblastoma cells by increasing protein levels of the oncosuppressor KCASH2" Oncogenesis 8(11):1-11 (2019).
  • Wang, D., Luo, H., Huo, Z., Chen, M., Han, Z., Hung, M., ... & Xiao, H. "Irradiation-induced dynamic changes of gene signatures reveal gain of metastatic ability in nasopharyngeal carcinoma" American Journal of Cancer Research 9(3):479 (2019).
  • Wen, M. H., Xie, X., Tub, J., Lee, D. F., & Chen, T. Y. "Generation of a genetically modified human embryonic stem cells expressing fluorescence tagged ATOX1" Stem Cell Research 101631: (2019).
  • Wilson, M. R., Reske, J. J., Holladay, J., Wilber, G. E., Rhodes, M., Koeman, J., ... & Patterson, A. L. "ARID1A and PI3-kinase pathway mutations in the endometrium drive epithelial transdifferentiation and collective invasion" Nature communications 10(1):1-18 (2019).
  • Yekula, A., Minciacchi, V. R., Morello, M., Shao, H., Park, Y., Zhang, X., ... & Carter, B. "Large and small extracellular vesicles released by glioma cells in vitro and in vivo" Journal of Extracellular Vesicles 9(1):1689784 (2019).