Mycoplasma Pro PCR Detection Kit
Cat. No.
G239
Unit
100 rxn
Price
$349.00
$249.00
| Cat. No. | G239 | ||||||||||||||
| Name | Mycoplasma Pro PCR Detection Kit | ||||||||||||||
| Description |
Mycoplasma Pro PCR Detection Kit is an improved version of our best-selling kit (Cat. No. G238) which now includes:
Timely detection of Mycoplasmas in cell cultures is recommended to deter wide-spread contamination and save on the costly efforts of elimination. This kit includes a PCR MasterMix containing gel loading dye for added convenience. Kit Features:
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. |
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| Unit | 100 rxn | ||||||||||||||
| Storage Condition |
Store at -20°C. This product is stable for 2 years from the date of shipping if stored and handled properly. |
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| 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. G239 |
| How do I use the Mycoplasma PCR Detection Kit in conjunction with the MycoAway™ Elimination Cocktail (1000X)? | |
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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).
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| Can I use this kit for detecting Mycoplasma in various cell culture supernatants? | |
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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.
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| Is there a specific protocol I need to follow when using the kit, or is it straightforward? | |
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We recommend utilizing the protocol listed in our datasheet since it was specifically developed for our Mycoplasma Pro PCR Detection Kit (Cat. No. G239). G239 will show a positive PCR amplification band (180 bp, internal control). Any sample that fails to amplify this is deemed inconclusive and testing should be repeated. Inhibition of PCR is possible if the cell culture sample has excessive debris or if too much sample is used. We recommend diluting the cell culture sample tenfold if the internal PCR control fails to amplify.
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| What are the specific advantages of using your kit compared to other commercially available Mycoplasma detection kits? | |
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abm’s Mycoplasma Pro PCR Detection Kit (Cat. No. G239) is rapid, comprehensive, and easy-to-use. Moreover, it was developed for more stringent cycling conditions and is capable of detecting additional Mycoplasma and Acholeplasma strains such as A. laidlawii. Finally, the product includes an internal PCR control which demonstrates that the PCR amplification was truly efficient and no inhibition or technical errorsoccured.
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| Are there any limitations to the types of Mycoplasma strains that can be detected with this kit? | |
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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.
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| Can you provide more details on the gel loading dye in the proprietary BlasTaq MasterMix formulation and how it facilitates gel electrophoresis? | |
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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.
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| 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? | |
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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? | |
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Non-specific amplifications can be a result of DNA contamination, excessive templates, and/or wrong annealing temperature. This can be prevented by touchdown PCR.
Touchdown PCR:
Lid 105°C Volume 25 μl
1) 95°C for 3min 2) 95°C for 15s 3) 70°C for 15s
4) 72°C for 15s
5) Go to Step 2, x3
6) 95°C for 15s 7) 65°C for 15s
8) 72°C for 15s
9) Go to Step 6, x3
10) 95°C for 15s 11) 60°C for 15s
12) 72°C for 15s
13) Go to Step 10, x25
14) 72°C for 1min 15) 4°C hold
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| What is the limit of detection of this kit? | |
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Our kit is able to detect as low as 10 copies of Mycoplasma/sample.
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| 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? | |
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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.
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| Will DMSO affect my PCR run? | |
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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.
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| I did not see amplification of the positive control. What can I try? | |
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This could occur in cases where the PCR machine does not ramp up temperature fast enough. We suggest trying touchdown PCR in these situations.
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| I did not see amplification of the internal PCR Band. What should I do? | |
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PCR amplification can be inhibited by excessive cells, debris, and/or byproducts. If PCR inhibition is noted and the 180 bp internal PCR control band is not seen, dilute the media sample with Nuclease-Free Water until the internal PCR control band is clearly seen (recommended dilution range 1/10 or 1/100).
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| My culture media contains antibiotics, will that be an issue? | |
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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.
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| We noticed that the band size for the positive control on agarose gel was around 400 bp, while the datasheet indicated it should be around 500 bp. Should we be concerned about this discrepancy? | |
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No, this is a normal observation. The positive control band should be strong and bright, with an expected size range between 400-500 bp on a well-resolved 2% gel. As long as the band is strong and clearly visible, it falls within the expected range, and there is no issue with the positive control.
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