Mycoplasma PCR Detection Kit
|Mycoplasma PCR Detection Kit
Enjoy superior performance at lower costs!
abm's 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 (Russell et al., 2020).
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.
Why test your cells for Mycoplasma?
Due to its small size, Mycoplasma can be present in your growth media without being visibly detected. This can cause significant errors in your experiments due to its effects on:
Many publishers now request Mycoplasma testing to be reported along with manuscript submissions to ensure accuracy in data reporting. Visit our learning resources center to read more about Mycoplasma contamination.
Store at -20°C. This product is stable for 2 years from the date of shipping if stored and handled properly.
|If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G238
Search CoA here
|What suggestions can you provide for the gel electrophoresis step?
Positive results will show a 370bp-550bp band so the PCR products can be ran on either a 1% or a 2% Agarose Gel with the proper corresponding DNA Markers. Please run the gel TAE Buffer with 100V for 20-25 minutes.
|Can samples be collected in advance and stored?
Collected samples can be stored at 4°C for 3-5 days before testing. 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.
|How do I use the Mycoplasma PCR Detection Kit in conjunction with the MycoAway™ Elimination Cocktail (1000X)?
Mycoplasma (Cat. No. G238) or Mycoplasma Pro (Cat. No. G239) Detection Kits should be used first in order to identify contaminated cells. Either product will only require a small volume (2.5 μl) of culture supernatant. If cells are Mycoplasma-positive, treat cells with MycoAway™ Elimination Cocktail (1000X) (Cat. No. G398).
|Can I use this kit for detecting Mycoplasma in various cell culture supernatants?
Yes, both Mycoplasma PCR Detection Kit (Cat. No. G238) and Mycoplasma Pro Detection Kit (Cat. No. G239) were tested in common laboratory media and serums.
|Is there a specific protocol I need to follow when using the kit, or is it straightforward?
We recommend utilizing the protocol listed in our datasheet since it was specifically developed for our Mycoplasma PCR Detection Kit (Cat. No. G238). If you encounter any challenge detecting Mycoplasma species, consider treating the samples with heat for 5 minutes at 95°C, cooling them down at 4°C, and adding to the reaction mixture.
|What are the specific advantages of using your kit compared to other commercially available Mycoplasma detection kits?
abm’s Mycoplasma PCR Detection Kit (Cat. No. G238) is rapid and comprehensive: more than 200 species will be detected in less than 2 hours. Furthermore, the product is easy-to-use with only 4 components present. Lastly, it is more affordable than other major competitors.
|Are there any limitations to the types of Mycoplasma strains that can be detected with this kit?
Our Mycoplasma PCR Detection Kit (Cat. No. G238) covers more than 200 species, while the Mycoplasma Pro PCR Detection Kit (Cat. No. G239) identifies additional Mycoplasma and Acholeplasma strains such as A. laidlawii. Please refer to our Mycoplasma Strains Guide listed under the Documents section in order to determine which detection kit satisfies your experimental needs.
|Can you provide more details on the gel loading dye in the proprietary BlasTaq MasterMix formulation and how it facilitates gel electrophoresis?
The MasterMix employs Bromophenol Blue sodium salt, a common dye in gel electrophoresis. As the compound is negatively charged, it migrates alongside the DNA making it possible to visualize the PCR product without the need to stain the gel.
|Can I expect consistent results over multiple uses, and how many tests can be conducted with a single kit? Can the collected supernatant be stored? If yes, what are the conditions for short-term and long-term storage?
The results will be consistent over multiple uses. The collected supernatant may be stored in case not all of the cell samples are ready at the same time. We recommend 4°C for short-term storage for up to 1 week and -80°C for long-term storage. Nevertheless, fresh cell culture samples will still provide more conclusive answers.
The kit is sufficient for 100 reactions.
|Gel electrophoresis shows non-specific amplifications. Any suggestions on how to prevent this?
Non-specific amplifications can be a result of DNA contamination, excessive templates, and/or wrong annealing temperature. This can be prevented by touchdown PCR.
Volume 25 μl
1) 95°C for 3min
2) 95°C for 15s
3) 65°C for 15s
4) 72°C for 15s
5) Go to Step 2, x3
6) 95°C for 15s
7) 60°C for 15s
8) 72°C for 15s
9) Go to Step 6, x3
10) 95°C for 15s
11) 55°C for 15s
12) 72°C for 15s
13) Go to Step 10, x25
14) 72°C for 1min
15) 4°C hold
|What is the limit of detection of this 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 detect the Mycoplasma?
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 2-3 days without changing the media so as to detect Mycoplasma in culture supernatant.
|Will DMSO affect my PCR run?
As long as the DMSO is diluted and the DMSO content is less than 0.5% in the PCR reaction it should not cause a problem. Too much DMSO would alter the Tm of the primers and thus affect 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 up temperature fast enough. We suggest trying touchdown PCR in these situations.
There are no references for this product yet!
Submitted on: 2023-12-13 21:21:25