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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
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For Use With (Equipment) |
Conventional PCR instrument |
|
Sample Type |
Strains, cell supernatants, etc. |
|
Polymerase |
Taq DNA Polymerase |
|
Detection Method |
Nested PCR |
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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 |
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Reaction time (fast PCR procedure) |
48 min |
Components
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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
Mycoplasma contamination is one of the most prevalent and insidious threats in cell culture, with estimates suggesting 15–35% of cell lines are contaminated at any given time. Unlike bacterial contamination, mycoplasma does not cause visible turbidity. Contaminated cells exhibit altered metabolism, gene expression, proliferation rates, cytokine secretion, and receptor signaling — invalidating experimental results. Regular testing is mandatory before publication, biobanking, or use in drug discovery assays.
This kit targets the most common cell culture contaminants, including Mycoplasma hyorhinis, M. orale, M. fermentans, M. arginini, M. hominis, Acholeplasma laidlawii, and others. Universal primer or probe designs used in PCR/qPCR-based kits detect conserved mycoplasma-specific sequences and may detect up to 70+ species. Refer to the species coverage table in the product datasheet for validated strains.
Best practices recommend testing all incoming cell stocks, after every 2–4 weeks in continuous culture, before cryopreservation, before experiments generating data for publication, and after any culture manipulation (transduction, transfection, co-culture). For GMP and biopharmaceutical environments, testing frequency and documentation must comply with regulatory agency guidelines (ICH Q5A/EMA/FDA).
Protocol requirements vary by kit format. MycAway colorimetric kits use culture supernatant directly (no extraction required), providing results in approximately 2 hours. PCR/qPCR-based kits (GMyc-PCR, qPCR variants) may require a brief extraction or heat lysis step for optimal sensitivity. The qPCR format provides quantitative data and is preferred for regulatory submissions. Refer to the specific product datasheet for the validated input protocol.
Yeasen Biotechnology offers flexible customization options for many of its assay kits and detection reagents, including custom lot sizes, bulk ordering, and application-specific formulation adjustments. Volume pricing, custom packaging, and kit bundling may be available depending on the product and intended workflow. A Certificate of Analysis (CoA) and lot-specific QC data are provided with every order. For inquiries regarding large-volume orders, custom configurations, or integration into automated workflows, please contact the BioHippo team for a tailored quotation.
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