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| Product Name | Mycoplasma PCR Detection Kit |
| Catalog No. | RESK-0189 |
| Description | Cell culture is a common experiment in life science research. Unlike other commonly used methods, cell culture is a dynamic and continuous process in which cells often respond to operational errors or contaminants with abnormal cell states or medium appearance. When contaminated with mycoplasma, however, cell morphology shows no obvious change, so contamination is easily overlooked and is often not discovered until it becomes severe. Several hundred mycoplasmas may adhere to the membrane of a contaminated cell; they compete for nutrients and release toxic metabolites, seriously affecting experimental results. Studies have shown that at least twenty mycoplasma species can contaminate cells, the most common being M. orale, M. arginini, M. hyorhinis, M. fermentans, M. hominis, M. salivarium, M. pulmonis, and M. pirum. The mycoplasma contamination rate of cultured cells ranges from 4% to 92%, with contamination sources including the working environment, the operators themselves (some mycoplasmas are part of the normal human flora), culture media, serum, cross-contamination between cells, laboratory equipment, and the original tissues or organs used to prepare the cells. This Mycoplasma PCR Detection Kit uses the PCR method to detect mycoplasma infection in various biological materials (such as cell cultures, laboratory animal secretions, and animal serum). It combines several advantages: sensitive, specific, rapid, and able to test cell culture supernatants directly. The primers are designed against the conserved region of the mycoplasma 16S-23S rRNA sequence and specifically amplify only mycoplasma DNA, achieving high detection sensitivity and specificity. PCR amplification and electrophoresis analysis take only a few hours, and the operation is convenient and simple. |
| Intended Use | For the detection of mycoplasma contamination in various biological materials, such as cell cultures, laboratory animal secretions, and animal serum. Good laboratory practice and regular testing for mycoplasma contamination are recommended, as any sudden change in cell culture should be regarded with suspicion. |
| Principle / Technology | Conventional single-round PCR. The primers are designed against the conserved region of the mycoplasma 16S-23S rRNA sequence and specifically amplify only mycoplasma DNA, followed by agarose gel electrophoresis analysis. |
| Detection Method | PCR amplification followed by agarose gel electrophoresis. |
| Sample Type | Cell culture supernatants (can be tested directly), cell suspensions (DNA extraction is required before PCR; the input volume is generally 1/10 of the reaction system or less), laboratory animal secretions, animal serum, and other biological materials. |
| Performance Range / Specifications | High sensitivity: mycoplasma at lower levels can also be detected, allowing earlier discovery of contamination. Strong specificity: when tested alongside other closely related bacterial samples, only mycoplasma is detected. Wide applicability: cell culture supernatants can be tested directly. Detects more than 20 common mycoplasma species. |
| Sensitivity / LOD | 10 copies/uL. |
| Specificity | No PCR amplification bands were observed for the following closely related organisms: Acinetobacter baumannii, Bacillus subtilis, Clostridium acetobutylicum, Clostridium perfringens, Klebsiella aerogenes, Lactobacillus acidophilus, Micrococcus luteus, Pseudomonas aeruginosa, Staphylococcus epidermidis, Streptococcus mutans, and Streptococcus pneumoniae. |
| Components / Formulation | Includes a positive control (Mycoplasma hyorhinis). |
| Storage Conditions | Store at -25 to -15 °C. If not used for an extended period, store protected from light. |
| Shelf Life | 18 months. |
| Package Specifications | 10 assays / 20 assays |
| Shipping Conditions | Shipped on dry ice. |
| Regulatory / Compliance | For research use only. |
| Application Notes / Precautions | If a cell suspension is used as the sample, DNA must be extracted before PCR. The input volume is generally 1/10 of the reaction system or less. Ethidium bromide (EB) staining: 1) If EB dye is added when preparing the agarose gel, it does not need to be added to the samples before loading; 2) If EB dye is not added to the agarose gel, the samples need to be mixed with EB dye before loading. The positive control is Mycoplasma hyorhinis. When comparing with one-step isothermal mycoplasma detection kits, note that the range of mycoplasma species covered by each method differs, so a sample positive by one method may be negative by the other if the contaminating species is outside the detection range. |
For research use only, not for clinical use.
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