Description
Mycoplasma PCR Detection Kit (Nested PCR) (2G) is based on the nested PCR method (Nested PCR) to detect mycoplasma infection in various biological materials (such as cell culture, experimental animal secretions and animal serum, etc.). The kit uses quantitative PCR technology, designs two sets of primers for the conservative region of the mycoplasma 16S-23S rRNA sequence, performs two rounds of PCR amplification, and judges the mycoplasma contamination based on the second round of amplification products. It can detect 34 types of mycoplasma, including oral mycoplasma (M.orale), pneumoniae (M.pneumoniae), hyorhinis (M.hyorhinis), arginine mycoplasma (M.arginini), gallisepticum (M.gallisepticum), synoviae (M.synoviae), fermentation mycoplasma (M.fermentans), citric spiroplasma (Spiroplasma citri), salivarium (M.salivarium), etc., with a sensitivity of up to 2.5 copies/uL. It can also amplify bands normally for 10 CFU/mL mycoplasma standard strains. It has the characteristics of strong specificity, high sensitivity, convenient and fast operation, and can be directly used for the detection of cell culture supernatant.
Compared with conventional PCR mycoplasma detection kits, this product can avoid the production of non-specific products due to non-specific pairing between primers and templates; it can also sensitively detect trace amounts of mycoplasma DNA in samples to avoid false negatives.
Features
- Comprehensive Detection: Advanced nested PCR technology enables the detection of up to 34 mycoplasma species in a single assay.
- Exceptional Specificity: Primers target the conserved 16S–23S rRNA region with no cross-reactivity to related species such as Clostridium or Lactobacillus.
- Ultra-High Sensitivity: Achieves a detection limit as low as 2.5 copies/μL for early and reliable identification.
- Fast & Convenient Workflow: Streamlined protocol allows for sample preparation and detection in under 2 hours.
- Safe & Contamination-Free: The included non-infectious positive control ensures safety and eliminates the risk of cross-contamination.
Specifications
|
For Use With (Equipment) |
Conventional PCR instrument |
|
Sample Type |
Strains, cell supernatants, etc. |
|
Polymerase |
Taq DNA Polymerase |
|
Detection Method |
Nested PCR |
|
Detection of mycoplasma types |
34 types, including 9 types of mycoplasma covered by the pharmacopoeia |
|
Detection limit |
1st Round PCR:1x103 copies/uL 2nd Round PCR: 2.5 copies/μL / 5CFU/mL |
|
Reaction time (fast PCR procedure) |
48 min |
Components
|
Components No. |
Name |
40614ES10 |
40614ES20 |
|
40614-A* |
GMyc-1st PCR Mix |
460 μL |
460 μL×2 tube |
|
40614-B* |
GMyc-2nd PCR Mix |
490 μL |
490 μL×2 tube |
|
40614-C** |
Positive Control Template |
20 μL |
40 μL |
【Note】: *PCR reaction is extremely sensitive. To prevent false positives, add the positive control last when adding samples.
**40614-C: Positive Control Template, positive control.
Shipping and Storage
This product should be stored at -25~-15℃ for one year.
*If the entire kit is not used for a long time, please store it away from light. If 40614-C is not used for a long time, it can be frozen at -85~-65℃.
Application
mycoplasma contamination; mycoplasma testing; Cell Culture Quality Control
Figures
1. Specificity
Figure 1. Specificity and matrix tolerance of GMyc-PCR Mycoplasma Detection Kit (2G)
(A)/(B).Sample matrix interference: 14 common sample matrices and the positive control and its spiked content (2.5 copies/μL positive control) and negative control (PCR Mix) in the kit were selected for verification. No mycoplasma was detected in any of the 14 matrices.
(C).Detection Range: GMyc-PCR Mycoplasma Detection Kit (2G) is capable of detecting all 8 mycoplasma species required by both Chinese and international pharmacopoeias, with a sensitivity of 10 CFU/mL.
2. High detection sensitivity
Figure 2. Sensitivity test
3. Stability
Figure 3. Stability Test
(A)Freeze-thaw stability: The kit withstands 10 freeze-thaw cycles at -25 to -15°C without loss of performance.
(B)Heat-accelerated stability: The Mycoplasma 2G nested PCR kit remains stable after 14 days at 37°C.
Citations & References:
[1] Rao XS, Cong XX, Gao XK, et al. AMPK-mediated phosphorylation enhances the auto-inhibition of TBC1D17 to promote Rab5-dependent glucose uptake. Cell Death Differ. 2021;28(12):3214-3234. doi:10.1038/s41418-021-00809-9(IF:15.828)
[2] Guo F, Li L, Li J, et al. Single-cell multi-omics sequencing of mouse early embryos and embryonic stem cells. Cell Res. 2017;27(8):967-988. doi:10.1038/cr.2017.82(IF:15.606)
[3] Hao Y, He B, Wu L, et al. Nuclear translocation of p85β promotes tumorigenesis of PIK3CA helical domain mutant cancer. Nat Commun. 2022;13(1):1974. Published 2022 Apr 13. doi:10.1038/s41467-022-29585-x(IF:14.919)
[4] Shu X, Liu M, Lu Z, et al. Genome-wide mapping reveals that deoxyuridine is enriched in the human centromeric DNA. Nat Chem Biol. 2018;14(7):680-687. doi:10.1038/s41589-018-0065-9(IF:13.843)
[5] Li X, Xiong X, Wang K, et al. Transcriptome-wide mapping reveals reversible and dynamic N(1)-methyladenosine methylome. Nat Chem Biol. 2016;12(5):311-316. doi:10.1038/nchembio.2040(IF:12.709)
[6] Sun L, Yang X, Huang X, et al. 2-Hydroxylation of Fatty Acids Represses Colorectal Tumorigenesis and Metastasis via the YAP Transcriptional Axis. Cancer Res. 2021;81(2):289-302. doi:10.1158/0008-5472.CAN-20-1517(IF:12.701)
[7] Sun Z, Zhang Z, Wang QQ, Liu JL. Combined Inactivation of CTPS1 and ATR Is Synthetically Lethal to MYC-Overexpressing Cancer Cells. Cancer Res. 2022;82(6):1013-1024. doi:10.1158/0008-5472.CAN-21-1707(IF:12.701)
[8] Song J, Zhuang Y, Zhu C, et al. Differential roles of human PUS10 in miRNA processing and tRNA pseudouridylation. Nat Chem Biol. 2020;16(2):160-169. doi:10.1038/s41589-019-0420-5(IF:12.154)
[9] He B, Pan H, Zheng F, et al. Long noncoding RNA LINC00930 promotes PFKFB3-mediated tumor glycolysis and cell proliferation in nasopharyngeal carcinoma. J Exp Clin Cancer Res. 2022;41(1):77. Published 2022 Feb 24. doi:10.1186/s13046-022-02282-9(IF:11.161)
[10] Tang B, Liu BH, Liu ZY, Luo MY, Shi XH, Pang DW. Quantum Dots with a Compact Amphiphilic Zwitterionic Coating. ACS Appl Mater Interfaces. 2022;14(24):28097-28104. doi:10.1021/acsami.2c04438(IF:9.229)
[11] Huang C, Zhang Z, Chen L, et al. Acetylation within the N- and C-Terminal Domains of Src Regulates Distinct Roles of STAT3-Mediated Tumorigenesis. Cancer Res. 2018;78(11):2825-2838. doi:10.1158/0008-5472.CAN-17-2314(IF:9.130)
[12] Wu X, Yu M, Zhang Z, et al. DDB2 regulates DNA replication through PCNA-independent degradation of CDT2. Cell Biosci. 2021;11(1):34. Published 2021 Feb 8. doi:10.1186/s13578-021-00540-5(IF:7.133)
[13] Wang J, Zhang Y, Liu X, Liu H. Optimizing Adaptive Therapy Based on the Reachability to Tumor Resistant Subpopulation. Cancers (Basel). 2021;13(21):5262. Published 2021 Oct 20. doi:10.3390/cancers13215262(IF:6.639)
[14] Feng W, Liu R, Xie X, et al. SUMOylation of α-tubulin is a novel modification regulating microtubule dynamics. J Mol Cell Biol. 2021;13(2):91-103. doi:10.1093/jmcb/mjaa076(IF:6.216)
[15] Yu M, Hu X, Yan J, Wang Y, Lu F, Chang J. RIOK2 Inhibitor NSC139021 Exerts Anti-Tumor Effects on Glioblastoma via Inducing Skp2-Mediated Cell Cycle Arrest and Apoptosis. Biomedicines. 2021;9(9):1244. Published 2021 Sep 17. doi:10.3390/biomedicines9091244(IF:6.081)
Related blog：
Mycoplasma Contamination Total Solution
Concept of Mycoplasma and the Impact of Contamination
Documents:
Safety Data Sheet
Manuals
40614_Manual_Ver.EN20250612.pdf
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