The positive control shows a band at 259 bp and depending on the concentration of mycoplasma DNA, an additional band at 481 bp due to the internal control. Internal controls should appear in every lane indicating a successfully performed PCR. This band may be less intense with increased amounts of amplicons formed, caused by
mycoplasma DNA loads of >5 ´ 104 copies/ml.
MP0035
LookOut® Mycoplasma PCR Detection Kit
Optimized for use with JumpStart™ Taq DNA Polymerase, D9307.
LookOut® Mycoplasma PCR Detection Kit
Synonym(s):
PCR-based mycoplasma screening
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1 KIT
$446.00
$446.00
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About This Item
NACRES:
NA.75
UNSPSC Code:
12352207
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Product Name
LookOut® Mycoplasma PCR Detection Kit, Optimized for use with JumpStart™ Taq DNA Polymerase, D9307.
quality
Not suitable for clinical diagnostic use. Will not detect clinically relevant species such as M. pneumoniae and U. urealyticum
packaging
pkg of 1 kit
technique(s)
PCR: suitable
application(s)
detection
microbiology
compatibility
Optimized for use with JumpStart™ Taq DNA Polymerase, D9307.
storage temp.
2-8°C
Quality Level
1 of 4
This Item | MP0040A | MP0030 | XNAPS |
|---|---|---|---|
| technique(s) PCR: suitable | technique(s) PCR: suitable | technique(s) PCR: suitable | technique(s) PCR: suitable |
| quality Not suitable for clinical diagnostic use. Will not detect clinically relevant species such as M. pneumoniae and U. urealyticum | quality Not suitable for clinical diagnostic use. | quality - | quality - |
| packaging pkg of 1 kit | packaging pkg of 1 kit | packaging pkg of 1 kit | packaging - |
mycoplasma detection, PCR-based assay | mycoplasma detection, qPCR-based assay | mycoplamsa removal agent, MRA, | XNAP, XNAPE, XNAPR, microbialanalysis |
| application(s) detection | application(s) detection | application(s) detection | application(s) - |
| compatibility Optimized for use with JumpStart™ Taq DNA Polymerase, D9307. | compatibility - | compatibility - | compatibility - |
Application
General description
The LookOut® Mycoplasma PCR Detection Kit utilizes the polymerase chain reaction (PCR), which is established as the method of choice for highest sensitivity in the detection of Mycoplasma, Acholeplasma, and Ureaplasma contamination in cell cultures and other cell culture derived biologicals. Kit has been successfully tested for the ability to amplify Mycoplasma fermentans genomic DNA in concentrations of 20,000 to 20 copies.
The reaction tubes included with the kit are pre-coated with appropriate dNTPs, primers, and loading dye. Total assay time is greatly reduced compared to general protocols that require individual loading of reaction tubes.
The reaction tubes included with the kit are pre-coated with appropriate dNTPs, primers, and loading dye. Total assay time is greatly reduced compared to general protocols that require individual loading of reaction tubes.
Other Notes
The kit has been optimized for use with Product No. D9307. Any DNA polymerase with a 3’-5’ exonuclease activity is NOT recommended. These include Tli, Pfu,and Pwo. Those DNA polymerases that may be used include Taq, Tfl, and Tth.
Legal Information
JumpStart is a trademark of Sigma-Aldrich Co. LLC
LookOut is a registered trademark of Merck KGaA, Darmstadt, Germany
related product
Product No.
Description
Pricing
Storage Class
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
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Articles
Detect mycoplasma contamination in cell culture through the PCR, DNA stain, or culture tests. Discover mycoplasma prevention, elimination, and detection kits.
Overview of good cell banking practices for cell line cryopreservation purposes.
Quality control guidelines to maintain high quality authenticated and contamination-free cell cultures. Free ECACC handbook download.
Overview on how to maintain proper cell culture laboratory safety including steps on performing lab risk assessments, surface disinfection, biohazard and waste removal. Free ECACC handbook download.
Protocols
Mycoplasma contamination of cell cultures is a serious issue impacting cell model validity. PCR testing for mycoplasma is an inexpensive, sensitive, and specific method for detecting contamination.
Related Content
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What could be the reason for multiple bands showing up in +control provided with kit (one strong band @ expected ~250bp and fainter band @ ~480-500bp - not expected)? Is there a way to fix this? All other samples run give single bands, including -control.
1 answer-
Helpful?
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What is the full comprehensive list of all Mycoplasma species that the LookOut® Mycoplasma PCR Detection Kit, not test for? I am aware of the two outlined species, M. pneumoniae and U. urealyticum, but I am not sure on how these two are clinically similar
1 answer-
This kit is intended for use as a mycoplasma screening and detection tool for cell culture use. It is not suitable for in vitro diagnostics use. While M. pneumoniae and U. urealyticum have been tested and found to be non-reactive, a full list of clinically associated mycoplasma species has not been determined. Please see the image below to review those species that have been tested using this kit.
Please use the following link to review the product Technical Bulletin:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/213/434/mp0035bul.pdfHelpful?
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What is the difference between product no. MP0040A (LookOut® Mycoplasma qPCR Detection Kit) and MP0035?
1 answer-
The main differences between these kits are as follows:
For MP0035:
•Standard PCR
•Primers/dNTPs precoated on tubes
•Includes tubes
•Taq Polymerase not included
•Requires 40 cycles at 30 secondsFor MP0040A:
•qPCR only
•Primers/dNTPs require addition to the reaction
•Does not include tubes
•Taq Polymerase included in primer/dNTP mix
•Requires 45 cycles at 30 seconds
MP0035 is suitable for 24 reactions, while MP0040A is suitable for 25 reactions.Helpful?
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What are the possible reasons for problems when trying to review results for MP0035-1KT?
1 answer-
Possible reasons for problems when trying to review results for MP0035-1KT could include incomplete removal of phenol during DNA preparation by phenol:chloroform extraction, leading to poor PCR results. It is also important to ensure the use of the recommended product, D9307, JumpStart™ Taq DNA Polymerase, as internal testing has indicated occasional sensitivity issues when using polymerases from other manufacturers. Additionally, the presence of inhibitors in the samples may affect the internal control band in the results. Lastly, mishandling of the vials and loss of lyophilized reagents could compromise the sensitivity of the assay.
Helpful?
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What are the components of the rehydration buffer for MP0035? Can the rehydration buffer M4694 be substituted by another buffer?
1 answer-
The rehydration buffer M4694 in Product No. MP0025 cannot be substituted by the PCR buffer P9367, and vice versa. The composition of the PCR 10X reaction buffer P9367 in the MP0025 kit is 100 mM Tris-Hcl, pH 8.5, 750 mM KCl, 30 mM MgCl2. Similarly, the replacement of Rehydration Buffer (Catalog Number M4694) with 10X PCR Buffer, Catalog No. P2192 is not possible. The composition of the 10× buffer P2192 is 100 mM Tris-HCl, pH 8.3 at 25°C; 500 mM KCl; 15 mM MgCl2; 0.01% gelatin.
Helpful?
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When should samples be heat inactivated for using MP0035, Mycoplasma PCR Detection Kit?
1 answer-
The possibility that as the slides dry, salts from the PBS are evaporating and leaving a white residue or film is considered.
Helpful?
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What is the required DNA concentration of the purified sample supposed to be to ensure compatibility with the LookOut® Mycoplasma PCR Detection Kit, MP0035?
1 answer-
The detection of Mycoplasma sp. with Product MP0035 - LookOut® Mycoplasma PCR Detection Kit requires less than 10 mycoplasma genomes per microliter of the sample, which is approximately 10 fg of DNA. It is important to ensure that the sample does not contain more than 100 µg/ml of DNA to limit non-specific amplification.
Helpful?
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Do mycoplasma-active antibiotics include pen/strep? Can media continue to be used with pen/strep prior to mycoplasma testing for kit R: MP0035?
1 answer-
Pen/strep can be used with the cell culture before assaying with this kit. Pen/strep does not affect mycoplasma, nor does it affect the sensitivity of this kit.
Helpful?
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What is the difference between product no. MP0040A (LookOut® Mycoplasma qPCR Detection Kit) and MP0035???
1 answer-
The key differences between these kit are as follows:
MP0035
Standard PCR
primers/dntps precoated on tubes
Tubes included
Taq Polymerase not included
Requires 40 cycles at 30 secondsMP0040A
qPCR only
primers/dntps require addition to reaction
Tubes not included
Taq Polymerase included in primer/dntp mix
Requires 45 cycles at 30 secondsMP0035 is suitable for 24 reactions while MP0040A is suitable for 25 reactions.
Please see the datasheets for both products to see experimental differences in the procedures:
MP0035
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/213/434/mp0035bul.pdf
MP0040A
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/109/594/mp0040abul.pdfHelpful?
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I purchased the JumpStart Taq DNA Polymerase in addition to the kit. This also included a 10x PCR buffer. Is this also needed or only the rehydration buffer of the kit?
1 answer-
This kit has been tested and optimized for use with JumpStart Taq DNA Polymerase, Catalog Number
D9307. The Jumpstart Taq 10X buffer supplied with the Jumpstart Taq is not used in this kit reaction set up. The Jumpstart taq is used alone with the rehydration buffer provided with the kit.Helpful?
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