PCR Mycoplasma Detection Kit
|Description||Mycoplasma detection is often difficult due to its lack of visable appearance, therefore it can be afflicting your valuable cell and affecting your results without your knowledge. Cells contaminated by mycoplasma species can have changes in proliferation, metabolism, gene synthesis and processing, and even adhesion properties. The solution for quick and easy mycoplasma detection is ABM's PCR Mycoplasma Detection Kit. The PCR Mycoplasma Detection Kit allows for fast and reliable identification of mycoplasma contamination in cell cultures. Mycoplasma DNA in the cell culture supernatant is amplified via PCR and visualized using gel electrophoresis. In addition to the short detection process (less than 2 hours), the easy handling and high sensitivity makes this PCR Mycoplasma Detection Kit a convenient tool for routine examination of cell cultures and media.|
|Unit quantity||100 Reactions|
This kit is PCR based and very easy to use. There is no initial training for that. But we will be more than happy to answer any questions related to the use of this kit.
Yes, we do have a positive control included in the kit.
We provide positive control in this kit as insurance for a positive PCR reaction.
We recommend culturing your cells in a 6-well plate or 10cm dish.
It is DNA fragments containing proprietary sequences diluted in medium.
That is normal as the contents are concentrated. You may warm the product in a 37 degrees Celsius water bath until the sample has completely dissolved.
Yes, this kit will also work for extracted DNA samples
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.
Our kit is able to detect as low as 10 copies of Mycoplasma /sample
You may use standard 1% agarose gel and this will be the most cost-effective option. The product size is small (less than 500 bp) so 1.5% to 2% gel can also be used.
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).
Storing the media samples at 4°C until you are able to centrifuge should be fine.
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.
We normally use 0.5X TAE buffer to cast 1% gel and also use the same buffer to run the electrophoresis in-house at 100V for about 20 minutes.
The Mycoplasma positive control is a cloned vector containing a section of Mycoplasma gDNA sequence.
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.
The qPCR mastermix should still be functional. The crystals are from our proprietary buffering salt, which will be dissolved eventually when you warm the solution up to room temperature or even 37C. All our qPCR mastermixes are very stable and can withstand this kind of warming cycle. Once the salt is fully dissolved, the vial can be kept at 4C for 2-3 months.
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.
We encourage customers to culture their cells in either a 6-well plate or 10 cm plate.
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