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MP0035 Sigma-Aldrich

LookOut® Mycoplasma PCR Detection Kit

Optimized for use with JumpStart Taq DNA Polymerase, D9307.

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Properties

Related Categories Assays & Reagents for Measuring Cytotoxicity, Proliferation & Viability Kits, Assays and Reagents for Measuring Cytotoxicity, Proliferation and Viability, Cell Analysis, Cell Biology, Cell Culture,
quality   Not suitable for clinical diagnostic use. Will not detect clinically relevant species such as M. pneumoniae and U. urealyticum
compatibility   Optimized for use with JumpStart Taq DNA Polymerase, D9307.
storage temp.   2-8°C

Description

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.

Application

Mycoplasma PCR Detection Kit Video Protocol

The LookOut® Mycoplasma PCR Detection Kit has been used for screening of mycoplasma infection.

Compatibility

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 Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis

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Certificate of Origin

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Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
What is the difference between Product No. MP0025 (VenorGem Mycoplasma Detection Kit) and Product No. MP0035 (Lookout Mycoplasma Detection Kit)?
The assay kits have a similar range of species detected (see product insert for listing). For Product No. MP0025, the components are supplied as individual reagents. For Product No. MP0035, the components are supplied in reaction tubes, already mixed and ready for addition of sample and Taq. The internal control for Product No. MP0035 is at 481 bp rather than 191 bp as found in Product No. MP0025.
How many tests can be performed with Product MP0035, LookOut® Mycoplasma PCR Detection Kit?
The kit contains 8 positive control reaction tubes and 24 test reaction tubes.A positive control and negative control should be run with each assay.Depending upon the number of samples run with each assay, the kit could test up to 24 samples.
Which species of Mycoplasma will Product MP0035, LookOut® Mycoplasma PCR Detection Kit, recognize?
Detection of all mycoplasma species tested so far and usually encountered as contaminants in cell cultures This kit will detect A. laidlawii, M. agalactiae, M. arginini , M. arthritidis, M. bovis, M. cloacale, M. falconis, M. faucium, M. fermentans, M. hominis, M. hyorhinis, M. hyosynoviae, M. opalescens, M. orale, M. primatum, M. pulmonis, M. salivarium, M. spermatophilum, M. timone. This kit is not suitable for detection of M. pneumoniae, U. urealyticum, or other clinically associated species.
How much DNA is necessary for use with Product MP0035, LookOut® Mycoplasma PCR Detection Kit?
Detection of Mycoplasma sp. requires less than 5 mycoplasma genomes per microliter of sample. This is approximately 1-5 fg of DNA.
When using Product MP0035, LookOut® Mycoplasma PCR Detection Kit, there is no band for the internal control?
When a customer observes a band with the positive control and no band for the negative control, it is possible that the Taq could be the problem. We observe a lack of sensitivity with some polymerases from other manufacturers. The activity of the polymerases was enough to amplify the positive control but not the internal amplification control. The internal amplification control is the meter for the sensitivity of the PCR and the activity of the Taq used. The positive control contains more DNA and is good for result evaluation, only. We recommend that the customer use Jump Start Taq (Product No. D9307) as indicated in the protocol.
Product MP0035, LookOut® Mycoplasma PCR Detection Kit does not contain TAQ. WhatDNA polymerase should be used?
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.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products.Once you log onto our website, you will find the price and availability displayed on the product detail page.You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 orview local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
My question is not addressed here, how can I contact Technical Service for assistance?
Ask a Scientist here.
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Cell Culture Guide
Protocols & Articles

Articles

Mycoplasma Detection and Elimination

Mycoplasma contamination is a major headache for the cell culturist. Mycoplasma are small, simple bacteria which lack a cell wall and represent one of the most prevalent and serious sources of cell l...
Edward Burnett and Liz Penn, European Collection of Cell Culture (ECACC®)
Don Finley, Market Segment Manager, MilliporeSigma, St. Louis, MO
Biofiles, Vol. 8, No. 18
Keywords: Amplification, Antibiotics, Cell culture, Eliminations, Metabolism, Polymerase chain reaction

Mycoplasma Detection/Elimination

The maintenance of contamination-free cell lines is essential to cell-based research. Among the biggest concerns are mycoplasma contamination. Although mycoplasma do not usually kill contaminated cel...
Don Finley
BioFiles v7 n1, 2011, 14–15
Keywords: Eliminations, Metabolism, Polymerase chain reaction

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Optimized PCR-based Detection of Mycoplasma [VIDEO]

The maintenance of contamination-free cell lines is essential to cell-based research. Among the biggest contaminant concerns are mycoplasma contamination. Although mycoplasma do not usually kill cont...
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Peer-Reviewed Papers
15

References

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