Specifications
| Description | Mycoplasma PCR Detection Kit (Cat. No. G238) offers highly specific and sensitive detection of 200+ strains of Mycoplasmas in less than 2 hours. This bestselling kit is designed to minimize false positives, ensuring quick and reliable routine screening of cell cultures. A recent official publication by a national public health agency found that our kit out-performs Sigma and Lonza’s kits while being 6X less expensive than ATCC’s kit. Mycoplasmas are highly undesirable, easily acquired, and notoriously difficult to detect, altering infected cells at the molecular level and leading to visible changes in cell morphology and growth characteristics. Timely detection of Mycoplasmas in cell cultures is recommended in order to deter wide-spread contamination and save on the costly efforts of elimination. Our proprietary MasterMix formulation contains a gel loading dye, making it convenient for gel electrophoresis.Kit Features:
Highly-sensitive PCR Mycoplasma Detection Kit has been designed and created with enhanced specificity for testing over 90 Mycoplasma, Acholeplasma, and Spiroplasma species in cell cultures with a 10 CFU/mL detection limit. It includes ready-to-use reagents which enable performing 20 reactions, each with internal control. The kit offers a PCR reaction mix specifically designed for mycoplasma detection, ensuring optimal performance. Its streamlined design enables simple and efficient discovery of mycoplasma contamination. With sample preparation taking just 10 minutes, you can obtain results within a few hours, allowing for swift and timely analysis. Features
This Mycoplasma PCR Detection Kit will be an ideal addition to your laboratory. It will give you the best final results and help you with sampling. It is fast, easy to use, and takes up less time. On top of that, it has controls and easy-to-interpret features. |
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| SKU | G238 |
| Storage Condition | Store at -20°C. This product is stable for 2 years from the date of shipping if stored and handled properly. |
| Unit quantity | 100 rxn |
Do you provide or sell a positive control so that we can be sure the assay worked?
Yes, we do have a positive control included in the kit.
What size plate, dish or flask should I use for the culturing of my cells?
We recommend culturing your cells in a 6-well plate or 10cm dish.
What is the basis of the positive control in the mycoplasma kit?
It is DNA fragments of Mycoplasma gDNA diluted in medium.
Can tissue culture supernatant be frozen and thawed for the test?
Yes.
I kept the cells in the presence of gentamycin, Should I use antibiotics-free media to detect mycoplasma in the media?
It is not necessary to remove routine antibiotics (e.g. penicillin, streptomycin, and gentamicin) from the media prior to PCR detection, as they will not interfere with the assay.
What is the detection limit of your kit?
Our kit is able to detect as low as 10 copies of Mycoplasma /sample
Our cells are grown in suspension and would not achieve 80% confluence, is there a target cell density (cells/ml) we should achieve to ensure we will detect the mycoplasma?
Mycoplasma testing is more dependent on how recently the cells have been subcultured. Seeding density does not play a major role when using this kit. We recommend cells be around at least 3-5 days old in culture without changing any fresh media so as to detect secreted mycoplasma in culture supernatant (i.e. the cell culture medium should not be refreshed/changed for 48-72 hours prior to collection).
Can I use cryogenically frozen supernatant?
Using samples stored at -80C should be fine for this kit. If you thawing out the cells and performing mycoplasma testing right away, usually when thawing the cells the supernatant from the vial is diluted when adding to the cell culture vessel. As long as the DMSO is diluted and the DMSO content is less than 0.5% final in the PCR reaction it should not cause a problem. Too much DMSO would alter the Tm of the PCR buffer and thus affect amplification.
Since I have such a large number of cell lines to test, can I store the cell culture supernatant prior to use in the PCR, or does it have to be freshly collected?
In general, the more fresh the sample is when running the PCR, the better; in keeping with this, it is normally advised to wait until all cell lines are ready to go prior to collecting the supernatant for each. However, if necessary, you can certainly store collected supernatant at +4C storage for short-term (1-2 days) storage; note that we highly recommend avoiding repeated freeze thaw cycles.
How many days after thawing my cells can I perform this mycoplasma test?
We recommend the cells should remain in culture for at least 48-72 hours prior to screening for the presence of mycoplasmas, and that the media sample collection only be done once the cells have reached at least 80% confluence.
There is a ~200-250bp band in one of my samples, does this mean it is mycoplasma positive?
In rare cases, there may be a weak non-specific binding to a region of human gDNA due to the degenerate nature of the primers that can amplify a weak 250bp band in human cell samples. Your sample can be considered as negative for mycoplasma, as mycoplasma amplicons will be 370-550bp in size. You can try to increase the annealing temperature of the PCR program to 60°C to reduce this non-specific amplification.
I did not see amplification of the positive control. What can I try?
This could occur in cases where the PCR machine does not ramp temperature fast enough. We suggest trying touchdown PCR in these situations.
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