EZ-PCR™ Mycoplasma Detection Kit

Highly-sensitive PCR assay designed with enhanced specificity to test for over 90 species of Mycoplasma, Acholeplasma, and Spiroplasma in cell cultures, with a detection limit of 10 CFU/mL. Includes ready-to-use reagents to perform 20 reactions, each with an internal control.
SKU: 20-700-20

Availability: In stock

$225.00

Description

Details

Product Overview:

The EZ-PCR™ Mycoplasma Detection Kit is a highly-sensitive and -specific PCR assay designed to test for the presence of over 90 species of Mycoplasma, Acholeplasma, and Spiroplasma in cell cultures with a detection limit of 10 CFU/mL. The kit contains a ready-to-use PCR reaction mix with highly-optimized mycoplasma-specific primers, a positive control, and an internal control, all for the simple and efficient discovery of mycoplasma contamination. Samples can be prepared in as little as 10 minutes and results can be easily obtained within just a few hours.

  • Easy to use. A complete PCR reaction mix that contains primers, Taq polymerase, and dNTPs is included.
  • Easy to run. Follow a simple protocol with ready-to-use reagents for as little as 10 minutes of preparation.
  • Easy to interpret. Internal and positive controls are included to ensure efficient and accurate PCR interpretation.


Avoid False Negatives - Internal Control Included

The EZ-PCR™ Mycoplasma Detection Kit includes an internal amplification control, a plasmid containing a non-mycoplasma-specific DNA sequence to test for false negatives. This internal control is simultaneously amplified in the tube with cell culture samples, and should be used with all PCR samples, including the negative and positive controls. This combination rules out inhibition from biological material, among other assay malfunctions that lead to misinterpretation of results, namely, false negatives.



 


Figure 1: Gel electrophoresis results obtained following PCR reaction preparation and amplification using the EZ-PCR™ Mycoplasma Detection Kit. Eight total reactions, including six samples, one negative control, and one positive control were tested, each of which contain the internal control to rule out PCR inhibition (i.e. false negatives). As shown, Sample 4 produced a mycoplasma-positive band at 270bp in addition to the internal control band at 357bp, while Samples 1, 2, 3, 5, and 6 produced only internal control bands and are thus negative for mycoplasma. Image courtesy of WiCell Research Institute.





The Importance of Routine Testing

Mycoplasma are one of the most common, yet elusive, contaminants of mammalian cell cultures. As the smallest known free-living organism, mycoplasma are a pervasive, parasitic species of highly-infectious bacteria that are estimated to contaminate between 15-35% of all continuous cell cultures worldwide. While other mycoplasma detection methods are available, PCR-based assays have the highest sensitivity with minimal preparation time for early detection in a rapid and simple manner, when compared to other methods.

The earlier mycoplasma contamination is discovered, the simpler it is to treat. The easy-to-use and cost-efficient EZ-PCR™ Mycoplasma Detection Kit was designed for highly-sensitive routine screening and detection of mycoplasma and other closely related species. To avoid major contamination, testing should be carried out minimally every 2 weeks to 3 months, especially when shared incubator spaces are used, as well as prior to the incorporation of new cultures from outside sources.

Efficient and precise PCR-based testing for mycoplasma infection should also be conducted throughout the cell culture manufacturing process from inoculation through harvest, involving routine tests of raw materials, cell banks, and viral seed stocks.

Figure 2. Images represent examples of mycoplasma that are capable of infecting all cell culture labs. Mycoplasma can be self-replicating and are the smallest known free-living prokaryote. Mycoplasma contamination can not be seen under a standard microscopes.


Requirements from Leading Scientific Journals and European Pharmacopoeia Guidance

Due to the prevalence of mycoplasma in cell cultures and, more importantly, the adverse effects on a number of cellular functions that contamination causes, many major, high-impact scientific journals have begun requiring proof of testing prior to publication. Moreover, regulatory guidance recommends that all products derived from mammalian cell culture be tested for the presence of mycoplasma. Already in 2007, the European Pharmacopoeia (5.8, Sec. 2.6.7) provided guidance on the validation requirements for nucleic acid amplification–based methods (PCR) as a substitute for the time-consuming, direct cell culture methods of detection.

Specifications

Specifications

QTY 20 reactions
Brand EZ-PCR™
Storage Conditions -20º
Shipping Conditions Dry Ice

References

references

  1. C.W.Q. Koh et al. Single-nucleotide-resolution sequencing of human N6-methyldeoxyadenosine reveals strand-asymmetric clusters associated with SSBP1 on the mitochondrial genome. Nucleic Acids Research, Volume 46, Issue 22, 14 December 2018, Pages 11659–11670
  2. A.Belostozky et al. Solidification of oil liquids by encapsulation within porous hollow silica microspheres of narrow size distribution for pharmaceutical and cosmetic applications. Materials Science and Engineering: C Volume 97, April 2019, Pages 760-767
  3. X. Bai et al. Establishment of an induced pluripotent stem cell line (FDEENTi001-A) from a patient with pathological myopia. Stem Cell Research, 2018
  4. E. Ruiz et al. Compositions for Protecting Skin Comprising DNA Repair Enzymes and Phycobiliprotein. US Patent App. 15/746,287, 2018
  5. M. Khoury et al. Treatment for Infection Composed of Menstrual Stem Cells US Patent App. 15/768,469, 2018
  6. A. Yoshimura et al. Exosomal miR-99a-5p is elevated in sera of ovarian cancer patients and promotes cancer cell invasion by increasing fibronectin and vitronectin expression in neighboring peritoneal mesothelial cells. BMC Cancer201818:1065
  7. K.L. Eales et al. Verteporfin selectively kills hypoxic glioma cells through iron-binding and increased production of reactive oxygen species. Scientific Reportsvolume 8, Article number: 14358 (2018)
  8. V. Alari et al. Generation of the Rubinstein-Taybi syndrome type 2 patient-derived induced pluripotent stem cell line (IAIi001-A) carrying the EP300 exon 23 stop mutation c.3829A > T, p.(Lys1277*), Stem Cell Research, Volume 30, July 2018, Pages 175-179

Documentation

Reviews

Customer Reviews (6)

Easy to use and fast Review by John-Mario Roussis
Quality
It's an easy and fast way to check whether your cell lines are contaminated with mycoplasma (Posted on 4/24/2018)
easy to use , very efficientReview by amina jbara
Quality
Easy to use, very efficient. (Posted on 1/15/2018)
very easy to useReview by Gidi Karmon
Quality
very easy to use, quick protocol, consistent results. (Posted on 1/11/2018)
ExellentReview by Orit Sagi-Assif
Quality
Very good product, get quick results.
The price is too high.
(Posted on 1/9/2018)
Good productReview by Sharon Tal
Quality
Very helpful. easy to use (Posted on 11/15/2017)
GoodReview by Gk
Quality
Good (Posted on 10/27/2017)

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