Mycoplasma qPCR Detection & Elimination Bundle

Cat. No.

G240-G398

Unit

Bundle

Price

$385.00

Cat. No.	G240-G398
Name	Mycoplasma qPCR Detection & Elimination Bundle
Description	Routine Monitoring Made Simple. Screen your cell cultures regularly with the fastest, most reliable qPCR kit. Inhibitor-Resistant MasterMix. Engineered to minimize inhibition and false negatives for confident results. Complete Protection. Detect early and eliminate effectively with one powerful bundle. abm's Mycoplasma qPCR Detection & Elimination Bundle combines the rapid sensitivity of our Mycoplasma qPCR Detection Kit (Cat. No. G240) with the powerful clearing action of MycoAway™ 1000X Mycoplasma Cocktail (Cat. No. G398) for complete mycoplasma management. The #1 qPCR Mycoplasma Detection Kit is optimized for routine monitoring, delivering ultra-sensitive detection of Mycoplasma and Acholeplasma directly from cell culture supernatant—no DNA extraction or gel electrophoresis required. Powered by our inhibitor-resistant 2X MegaFi™ Pro qPCR MasterMix, it minimizes inhibition and false negatives while detecting as few as 10 copies and eliminating NTC signals, all within 2 hours. The kit includes a separate ROX Reference Dye for broad qPCR instrument compatibility. Paired with MycoAway™ 1000X Mycoplasma Cocktail, which eradicates 70+ Mycoplasma species in just 2–3 weeks (4–6 passages) without harming your cells, this bundle offers both early detection and effective elimination—ensuring reliable quality control and long-term protection for your valuable cell lines. Not sure whether this kit detects your target species? Visit our Mycoplasma Kit Finder to confirm coverage and find the right kit.
Unit	Bundle
Material Citation	If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G240-G398

Datasheet

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	Is this kit compatible with all real-time PCR machines?
	The qPCR MasterMix contains dye comparable to SYBR Green™ and EvaGreen™. This makes the kit compatible with all standard SYBR-capable qPCR systems. ROX Reference Dye is provided separate from the MasterMix, making this kit universally compatible with most qPCR instruments. For specific instrument requirements, please refer to the Rox Machine Compatibility Chart.

	Are there any additional sample preparations required before running the qPCR?
	No additional sample preparation is required. Simply dilute your cell culture supernatant sample 1:5 using the provided Nuclease-free water prior to adding it to the qPCR reaction. This helps minimize inhibitors which could affect qPCR sensitivity.

	Can I use this kit for detecting Mycoplasma in various cell culture supernatants?
	Yes, this kit has been tested with common laboratory media and serums.

	The melt peak in my sample is at a different temperature than the positive control. Are my results still valid?
	Yes, your results can still be accurate. Because the primers are designed to detect a broad range of mycoplasma species and strains, the resulting amplicons can have slightly different sequences and lengths, which lead to variation in melt peak temperatures.

What if my NTC shows amplification?

Amplification in the NTC typically indicates:

Cross-contamination during setup
Reagent contamination

The following are suggested to avoid cross-contamination:

Include a no-template control (NTC) on every plate to monitor contamination. Ideally NTCs should be set up first prior to sample handling.
Ensure positive control material is handled last and kept physically separate from test samples. If possible, positive control should be set up in a separate room.
Always UV-sanitize lab and wipe down work surfaces and pipettes regularly with 10% Bleach.
Use filtered, aerosol-resistant pipette tips and change tips between every sample and reagent.
If unexpected amplification occurs in negative controls, repeat the run with freshly prepared reagents and clean equipment.

What is the limit of detection (LoD) of this kit?

This kit reliably detects as few as 10 copies of mycoplasma DNA per reaction. This is based on validation data showing consistent amplification up to Ct 35. Signals beyond Ct 35 are considered below the detection threshold and interpreted as negative.
If you suspect low-level contamination, we recommend continuing to culture the cells for 48–72 hours and re-testing, as the mycoplasma population may be below the detectable limit in early infections.

What are the species and strains detected using Mycoplasma qPCR Detection Kit (Cat. No. G240)?

The kit covers more than 300 species and strains of Mycoplasma including the most common species M. arginini, M. pneumoniae, M. hyorhinis, M. orale, and Acholeplasma laidlawii. Please refer to our Mycoplasma Strains Guide in order to determine which detection kit satisfies your experimental needs. If you cannot find the particular species or strain on the list, you can also email us at technical@abmgood.com and we can manually verify if the kit would detect your particular strain.

	Can samples be collected in advance and stored?
	Collected samples can be stored at 4°C for 3-5 days before testing, or -20°C if planning to store for a longer period of time. Please note, samples should be drawn from cells that have been in culture for at least 48-72 hours un-disturbed and are at least 80% confluent.

	Our cells are grown in suspension and would not achieve 80% confluence, is there a target cell density we should achieve to ensure Mycoplasma is detected?
	Mycoplasma testing is more dependent on how long the cells have been in culture. Seeding density does not play a major role when using the detection kit. We recommend cells be kept in culture for 48-72 hours without changing the media so as to detect Mycoplasma in culture supernatant.

	My culture media contains antibiotics, will that be an issue?
	It is fine to test supernatant containing antibiotics. Please ensure that the culture has been in incubation for at least 48-72 hours prior to sample collection.

	What type of polymerase is used in this mycoplasma detection kit?
	MegaFi™ Pro 2X qPCR MasterMix uses a Type B family DNA polymerase, which is optimized for reliable and accurate amplification in qPCR. Unlike Taq polymerase, it does not produce 3’-A overhangs, which helps reduce background and improves the clarity of results in detection workflows.

	What is the positive control made of?
	The positive control contains a Mycoplasma genomic fragment cloned into a vector.

	Can the primers be used with other qPCR Mastermixes?
	No. It is recommended to only use the component included with the kit. Use of other mastermixes is not recommended and cannot be guaranteed to perform reliably, as sensitivity and specificity may be compromised.

Can I store culture supernatant before running PCR/qPCR?

Yes. If you can’t run PCR/qPCR right away, you can store the supernatant using proper conditions to protect DNA.

Storage guidelines:

Up to 24 hours: Store at 4 °C
Longer storage: Aliquot first, then freeze
- –20 °C: Good for weeks to a few months
- –80 °C: Best for long-term DNA stability

Important tips:

Use clean, nuclease-free tubes
Label clearly (sample ID + date)
Avoid repeated freeze–thaw cycles — this can reduce PCR/qPCR sensitivity
If you’ll store samples regularly, validate with control samples to make sure results stay consistent

	Is this kit validated for GMP or pharmacopeia mycoplasma testing?
	No. This kit is not validated to USP, EP, or cGMP guidelines for mycoplasma detection and is not certified specifically for GMP/GLP use. The manufacturer holds MDSAP certification, which reflects quality system compliance but does not replace pharmacopeia or GMP validation requirements.

Why can this assay detect mycoplasma without a DNA extraction step?

Mycoplasma lack a cell wall and are found mainly in the culture supernatant of contaminated cells. After 48–72 hours of incubation, enough mycoplasma and their DNA accumulate in the supernatant to allow direct PCR detection. The kit uses a highly sensitive, inhibitor-tolerant PCR mix that can detect very low DNA copy numbers, eliminating the need for a separate DNA extraction step and reducing hands-on time and contamination risk.

	What are the limitations of this direct PCR method?
	This method is best for routine testing of properly incubated cultures (48–72 hours). Very early or low-level contamination may not be detected without sufficient incubation. The assay detects mycoplasma DNA only and cannot distinguish between live and dead organisms.