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Cat. No.: MCLK-25 (for 25T)

Cat. No.: MCLK-100 (for 100T)

Description

GoodColor™ Mycoplasma Red and Yellow Colorimetric LAMP Assay Kit with UDG) amplifies mycoplasma DNA via Loop-Mediated Isothermal Amplification (LAMP) and identifies mycoplasma contamination in cultured cells and other biological samples relying on sensitive red-to-yellow color transition. It delivers superior analytical sensitivity and clearer positive/negative color discrimination versus the GoodColor™ Mycoplasma Colorimetric LAMP Assay Kit. Integrated with dUTP incorporation plus UDG-mediated amplicon carryover prevention technology, the kit enables rapid, high-sensitivity mycoplasma screening with visual readout; no agarose gel electrophoresis is required, and results are interpreted visually by naked-eye color change.

Mycoplasmas are the smallest and simplest prokaryotes. Key biological traits are listed below: devoid of cell walls, mycoplasma are inherently resistant to cell-wall-targeted antibiotics such as penicillin and other β‑lactams. Measuring 0.2–0.8 μm in diameter (intermediate between bacteria and viruses), several mycoplasma species pass through standard 0.22 μm sterile filters, rendering routine sterile filtration ineffective for mycoplasma removal. Limited intrinsic biosynthetic pathways force mycoplasma to source essential nutrients from host cells, enabling them to adhere to or scatter across cell surfaces and intercellular spaces. These physiological characteristics make mycoplasma contamination a prevalent global challenge in cell culture workflows.

Mycoplasma contamination severely compromises cellular physiology by altering host gene expression and metabolic profiles, triggering retarded proliferation, aberrant differentiation or unexpected cell death, and impairing overall cellular functionality. Such artifacts drastically undermine experimental reproducibility, consistency and data reliability, highlighting the necessity of routine mycoplasma surveillance.

Bacterial, yeast or fungal contaminants in cell culture are readily visualized under light microscopy, whereas mycoplasma cannot be detected via conventional optical observation and demand dedicated analytical assays. Established mycoplasma testing workflows include microbial culture isolation, ELISA, bioluminescence assays, nested conventional PCR, qPCR and fluorescent DNA staining. Limitations exist across these classic methods: mycoplasma isolation is labor-intensive; ELISA suffers from insufficient sensitivity and specificity; standard nested PCR is time-consuming and requires post-amplification tube opening for electrophoresis; qPCR and luminescence detection rely on specialized laboratory equipment. In contrast, LAMP proceeds under constant isothermal incubation and generates colorimetric readouts without downstream electrophoresis.

Isothermal amplification is an in vitro nucleic acid amplification technique performed at a fixed constant temperature, where combinatorial enzymes and target-specific primers drive rapid nucleic acid amplification. This kit adopts Loop-Mediated Isothermal Amplification (LAMP): 4–6 tailored primers bind distinct conserved regions of the target gene, and strand-displacement Bst DNA Polymerase initiates DNA synthesis to form dumbbell-shaped amplicons that fuel continuous cyclic strand displacement amplification. Final LAMP products consist of a heterogeneous pool of cauliflower-like DNAs with variable stem-loop structures. Complete amplification finishes within 30–60 min under steady isothermal incubation (60–65 °C). Unlike standard PCR, LAMP eliminates repetitive thermal denaturation cycles, gel electrophoresis and UV imaging, featuring simplified operation, rapid turnaround, outstanding sensitivity and robust target specificity.

This kit is equipped with anti‑carryover contamination technology to effectively avoid amplicon contamination. Formulated with dUTP and thermolabile UDG, the reagent enables dU incorporation into amplified products. Pre‑incubation at 37 °C for 5–15 min before isothermal amplification efficiently eliminates carryover contamination from prior amplicons. The thermolabile UDG gets inactivated during LAMP incubation and causes no interference to subsequent isothermal amplification.

Supplied with Myco Positive Control for validating kit functionality, this control material contains no mycoplasma and poses zero contamination risk. Running positive control alongside every batch of testing is recommended.

For standard 20 μL reaction setup, the small pack supports 25 tests and the medium pack enables 100 tests.

Components

25T

• LAMP Master Mix with UDG (2X)    250μl
• Bst DNA Polymerase    25μl
• Myco Positive Control    50μl
• Nuclease-free Water    200μl
• Mineral Oil    500μl

• LAMP Master Mix with UDG (2X)    1ml
• Bst DNA Polymerase    100μl
• Myco Positive Control    200μl
• Nuclease-free Water    1ml
• Mineral Oil    2ml

Features

High Sensitivity & Short Assay Runtime: The kit exhibits roughly 10-fold higher sensitivity compared to conventional PCR, with a calculated limit of detection of 20–200 copies/μl quantified using positive control plasmids. Total hands-off incubation finishes within one hour.

Excellent Specificity & Broad Mycoplasma Coverage: Primers are designed against conserved 16S rRNA genomic sequences of mycoplasma, permitting direct template testing using cell culture supernatant, serum and other biological specimens. The kit detects approximately 45 mycoplasma species, covering all common cell-culture contaminants, with no cross-amplification against bacterial, fungal or mammalian eukaryotic genomic DNA.

Visual Colorimetric Endpoint, Electrophoresis-Free: Formulated with optimized red chromogenic indicator dye, the assay relies on straightforward visual interpretation: robust target amplification shifts reaction color from crimson/rose red to orange-yellow / bright yellow, confirming positive mycoplasma contamination. Compared to GoodColor™ Mycoplasma Colorimetric LAMP Assay Kit, this formulation yields more stable color transition and markedly sharper differentiation between positive and negative reactions.

Storage

Store at -20°C, valid for one year. The LAMP Master Mix with UDG (2X) should be stored away from light. Avoid repeated freeze-thaw cycles as much as possible.

Precaution

• Before use, ensure that the reagents are completely thawed and gently mix by inverting the tubes. Try to avoid bubble formation during the mixing process.
• Based on testing, this product has shown no significant impact on performance after undergoing 10 freeze-thaw cycles. However, it's still advisable to minimize repeated freeze-thaw cycles, as they might cause a decrease in product performance.
• Isothermal amplification is an extremely sensitive detection method, so it's recommended to conduct testing in a standard PCR laboratory. The setup for isothermal amplification reactions should be performed in an area that minimizes the risk of contamination from various potential amplification products. Although this assay kit uses UDG enzyme for contamination prevention, please do not open the PCR tube cap. After isothermal amplification, seal the amplification products as required to prevent highly concentrated amplification products from contaminating the laboratory environment due to aerosols and other factors.
• This product contains a red indicator. Prior to the reaction, if the color after sample addition differs markedly from the negative and positive control colors, appropriately dilute the sample, pretreat it with a DNA extraction kit, or adjust the sample pH to approximately 8.0. Do not use nucleic acid preservation solutions containing Tris or other buffering agents; ultrapure water or double-distilled water is recommended for nucleic acid storage.
• While this assay kit can detect common Mycoplasma species responsible for cell contamination, for important cells suspected of potential contamination by Mycoplasma species that cannot be identified, it is recommended to use two or more methods based on different principles to detect simultaneously, effectively avoiding false positives and false negatives.
• This isothermal amplification system requires high concentrations of divalent ions; the sample must not contain high concentrations of metal ion chelators like EDTA.
• It is recommended to use filter-tipped pipette tips to set up the isothermal amplification system, which can maximize the prevention of contamination-induced false positives.
• This product is intended for scientific research by professionals only. It must not be used for clinical diagnosis or treatment, food or drug purposes, and should not be stored in residential settings.
• For your safety and health, please wear lab coats and disposable gloves during operation.

Only for research and not intended for treatment of humans or animals

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