Mycoplasma Detection and Elimination Reagents

  • Mycoplasma Growth Media

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    PurMa™ Mycoplasma Growth Media

    PurMa™ Mycoplasma Growth Media is a proprietary combination which provides an excellent environment for various species of mycoplasma to grow.

    Nutrition

    Mycoplasma has a complex nutritional requirement. This therefore, makes them dependent on external supplies. PurMa™ Mycoplasma Growth Media is ideal for growing different strains. Mycoplasmas have three important features: they are categorized as Mollicutes class, they also possess low GC content, and they basically don’t have cell wall. Additionally, mycoplasma has a simplified metabolic pathway and therefore it is a hard to grow this organism, especially under laboratory conditions.

    Mycoplasma Detection

    PurMa™ Mycoplasma Growth Media is also used for detecting the exact strain of contaminating mycoplasma. The best tool for evaluating  the presence of mycoplasma in any liquid such as media is by using the PCR method. The mycoplasma agar bcan also be used by growing mycoplasma and counting mycoplasma fried egg” shape colonies.
    • Initially,  Incubate the contaminated samples of media, FBS or cell line in 5 ml of PurMa™ MycoGrowth media (liquid).  The condition must be anaerobic while shaking.
    • Pipet 100 µl of the media on an PurMa™ MycoAgar plate.
    • After three days individual fried egg structure will start to show up.
    • Pick up several colonies and grow them in PurMa™ MycoGrowth liquid media.
    • Use PurMa™ Mycoplasma PCR Detection kit (Cat#: P5M011404) to amplify the fragment.
    • Have the PCR reaction cleaned and sent for sequencing.
    • The 230bp PCR fragment resulted from our kit will clearly determine the strain of your contaminant.

    Special Features

    • Easy -to- Use
    • Is an excellent tool to determine your exact mycoplasma contamination.
    • Contains a proprietary formulation which maximizes the growth of all known mycoplasmas.
    • Rigorously tested for bacteria, endotoxin, and mold.
    • Free of all mycoplasmas inhibiting growth elements.

    References

    1. DEFINED MEDIUM FOR MYCOPLASMA LAIDLAWII. TOURTELLOTTE et al. J Bacteriol. 1964 Jul;88(1):11-5.
    2. Comparison of undefined medium and its dialyzable fraction for growth of Mycoplasma. Pollock et al. 1969 Feb;97(2):522-5.
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    Parameter Specification
    Appearance Liquid/Powder
    pH 7.4 ± 0.1
    Osmolality 275-360 mOsm/L
    Endotoxin NMT< 2EU/mL
    Mycoplasma Negative
    Suitability  Suitable for mammalian cell culture
    Indicator NA
    Sterility Tested Sterile filtered using 0.22 µm filter
    Form Liquid/ powder
    Shipping Condition Ambient
    [/fusion_table][/fusion_builder_column][/fusion_builder_row][/fusion_builder_container]500 g, 1 kg, 1 x 500 ml, 1 x 1000 ml Read more

  • Mycoplasma PCR Detection Kit

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    Mycoplasma PCR Detection Kit

    Background

    Undoubtedly, PurMa Biologics is the pioneering company in both mycoplasma detection and elimination reagents. Moreover, PurMa™ Mycoplasma PCR Detection Kit is fast, easy to use, highly accurate kit. The kit contains a 2X green master mix and needs just 1-2µl of your sample to be tested.

    PurMa™ Mycoplasma PCR Detection Kit (50 Reactions) Cat # P5M011404-50 Contents

    • PurMaTM PCR 2X Mix (containing green dye, no need for loading dye  includes all the primers) (700 µl)
    • It also contains reaction buffer (2500µl)
    • More importantly, Positive control. The lysed mycoplasma which does not carry any live mycoplasma (50ul)
    • Negative Control. Lysed COS1 cells serves as the negative control (mycoplasma free), ready to proceed to the PCR and (50µl).
    • Lastly, PurMa™ Ultra-Pure Nuclease Free Water (Cat# P3W013104) (1ml)
    To gain the best results, please strictly follow the guidance: Protocol

    Mycoplasma Contamination

    Contamination with mycoplasma does not cause turbidity or obvious changes in cell culture media, therefore is under-looked in many laboratories. Although, to date, more than 190 mycoplasma species have been reported, it is eight species that account for more than 95% of all mycoplasma contamination in cell culture, including: M. arginini (bovine), M. fermentans (human), M. hominis (human), M. hyorhinis (porcine), M. orale (human), M. pirum (human), M. salivarium (human), and Acholeplasma laidlawii (bovine). Our sophisticated PurMa™ Mycoplasma PCR Detection Kit not only detects the mycoplasma in your cell culture system, but also determines the exact mycoplasma strain is the cause of contamination.

    Consequences of contamination with mycoplasma

    1. Gradual deterioration of the cell line, which is especially significant in stable cell lines (15-35%)
    2. Inconsistency among the results and much higher standard deviations in your statistical analysis.
    3. Altering DNA, RNA, and protein synthesis.
    4. Diminishing amino acid and ATP levels.
    5. Chromosomal alterations.
    6. Modifying host-cell immune system.

    Mycoplasma contamination can happen in various ways

    1. Faulty laboratory equipment. (For Instance: a dysfunctioning laminar flow could let mycoplasma-containing dust in the biosafety cabinet, thereby causing contamination of all of the media and cells within.)
    2. Inadequate culturing techniques, human breath contains several major mycoplasma including M. fermentans which count for the majority of cell culture contamination (45%). %). Wearing a mask is absolutely needed while working in a cell culture lab, especially when checking cells under microscope.
    3. Getting a cell line from another laboratory may introduce Mycoplasma contamination.

    Mycoplasma can affect the phenotypic and functional characteristics of cells In Vitro

    Including:
    • Chromosomal aberrations
    • Disruption of nucleic acid synthesis
    • Changes in membrane antigenicity
    • Inhibition of cell proliferation and metabolism
    • Decreased transfection rates
    • Changes in gene expression profiles
    • Affects virus production.
    • Cell death

    Special Features

    • PurMaTM Mycoplasma PCR Detection Kit is Easy to Use
    • The procedure is fast and is completed within 2 hrs.
    • Our kit has high detection ability.
    • Does not detect bacterial or animal sequences of DNA.
    • PurMaTM Mycoplasma PCR Detection Kit detects the following species:  A. Laidlawii, M. Arginine, M. Boris, M. Fermentans M. Hominis, M. Hyorhinis and M. Orale
    • PurMaTM Mycoplasma Detection Kit detects a 230 bp amplicon.
    • The kit contains a 2X ready master mix containing all the primers and green dye (you would not need to add loading dye after amplification to tun the result on agarose gel) and needs only 1-2µl of your sample to be tested.

    References

    1. Mycoplasma detection in cell culture by concomitant use of bisbenzamide and fluoresceinated antibody. Freiberg et al. In Vitro Cell Dev Biol. 1990 Jun;26(6):585-8.
    2. Mycoplasma detection by PCR analysis. Hopert et al. In Vitro Cell Dev Biol Anim. 1993 Oct;29A(10):819-21.
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    Parameter Specification
    Appearance Clear liquid/Green
    pH 7.4 ± 0.1
    Osmolality 275-360 mOsm/L
    Endotoxin NMT< 2EU/mL
    Mycoplasma Negative
    Suitability  Suitable for mammalian cell culture
    Indicator NA
    Sterility Tested Sterile filtered using 0.1 µm filter
    Form Liquid
    Shipping Condition Dry Ice
    [/fusion_table][/fusion_builder_column][/fusion_builder_row][/fusion_builder_container] Read more

  • Mycoplasma Removal Spray

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    PurMa™ Mycospray

    Background

    PurMa™ Mycospray is the most sophisticated Mycoplasma removal agent currently available in the market. Currently, an estimated 25% of U.S. laboratory cell cultures are contaminated with various species of mycoplasma.

    Source of Mycoplasma to your laboratory

    1. The lack of HEPA filter and positive pressure. Without the mentioned condition, there is certainly a flow of mycoplasma in the lab.
    2. Moreover, lab personnel is the second main source of mycoplasma contamination.
    So, entering mycoplasma to your tissue culture room is inevitable. As a result,  the tissue culture hood, CO2 incubator, benches, pipettors and the gloves you wear need to be routinely treated by a mycoplasma disinfectant.

    PurMa™ Mycospray Special Features

    • PurMa™ Mycospray is formulated to destroy the mycoplasma nuclear composition and will not leave any mycoplasma if it is used as instructed below.
    • Moreover, PurMa™ Mycospray is safe for skin and does not create any vapor to inhale at room temperature.
    • PurMa™ Mycospray destroys the following species with 100% certainty: A. laidlawii, M. arginine, M. boris, M. fermentans M. hominis, M. hyorhinis and M. orale.

    Instruction to use

    1. Because of the massive flow of mycoplasma to your tissue culture room, PurMa™ Mycospray needs to be used routinely at least three times per week.
    2. After completing work in tissue culture lab, routinely, wipe inside the hood and all the surfaces including the microscope area where you view your cells as well as the pipets. This is however, applies to other instruments which you do not keep under the UV light.
    3. Apply PurMa™ Mycospray , DO NOT dry it. It is therefore, crucial to leave the surface wet as it takes 5 minutes to complete the destruction of mycoplasma nuclei membrane.
    4. On the following day clean the mentioned surfaces with 70% alcohols to remove the residues of the dead mycoplasmas.
    5. Wear mask at all times inside the tissue culture lab as breathing is one of the main sources of several mycoplasma including m. Fermentans m. Hominis, m. Hyorhinis and m. Orale
    6. Wash your gloves with PurMatm Mycospray before any cell culture work

    References

    1. Bioprocess: Robustness with Respect to Mycoplasma Species. Faison et al. PDA J Pharm Sci Technol. 2020 Mar-Apr;74(2):201-212. Epub 2019 Sep 13.
    2. Mycoplasma removal: simple curative methods for viral supernatants. Baronti Cet al. J Virol Methods. 2013 Feb;187(2):234-7.. Epub 2012 Oct 29.
    [/fusion_text][fusion_table fusion_table_type="1" fusion_table_rows="2" fusion_table_columns="2" margin_top="" margin_right="" margin_bottom="" margin_left="" hide_on_mobile="small-visibility,medium-visibility,large-visibility" class="" id="" animation_type="" animation_direction="left" animation_color="" hue="" saturation="" lightness="" alpha="" animation_speed="0.3" animation_delay="0" animation_offset=""]
    Parameter Specification
    Appearance Clear  liquid
    pH 8.6 ± 0.1
    Osmolality 275-360 mOsm/L
    Endotoxin NMT< 2EU/mL
    Mycoplasma Negative
    Suitability Suitable for mammalian cell culture NOT to be in contact with Cells
    Indicator NA
    Sterility Tested Sterile filtered using 0.1 µm filter
    Form Liquid
    Shipping Condition Ambient
    [/fusion_table][/fusion_builder_column][/fusion_builder_row][/fusion_builder_container]500 ml spray (1X), 500 ml (10X), 1 Liter (10X) Read more

  • Mycoplasma Treatment Kit (MTK) (100X)

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    PurMa™ Mycoplasma Treatment Kit (MTK) (100X)

    PurMa™ Mycoplasma Treatment Kit (MTK) (100X) is a valuable tool for eliminating mycoplasma from live cells.

    Important Note

    Completing the eradication of mycoplasma from a contaminated cell line is laborious, time consuming as well as expensive. The procedure might also cause more contamination. So, we strongly recommend disposing of the contaminated cell line and starting a new culture if possible. This, however, might not be possible due to high value cell lines such as monoclonal antibodies or stable cell lines.

    Feeder Cells

    During the eradication process, fresh spleen cells of a healthy mouse are needed. Before starting the process: Isolate the spleen cells from a healthy adult mouse. Resuspend in 10 ml media of choice containing 20% FBS in a 15ml snap tube and place in 37°C CO2 incubator. For detailed procedure regarding the isolating spleen cells, click here: Detailed Procedure

    before you start

    1. Decontaminate your CO2 incubator, Hood, and benches before starting the procedure, otherwise in no time, the contamination will be back into your cultured cells.
    2. Use unopened media bottles and fresh FBS.
    3. During the following mentioned procedure and after that, wear mask at all times while you work in cell culture lab.
    4. Spray your gloves, bench, hood with PurMa™ Mycospray (1X) (Cat#: P5M010405) before and after work. PurMaTM Mycospray (1X) instantly eliminates/ deactivates mycoplasma.
    For detailed protocol of mycoplasma elimination, Click here: PurMa™ Complete Mycoplasma Removal for Cell Culture

    References

    1. Eradication of mycoplasma contaminations. Uphoff et al. Methods Mol Biol. 2013;946:15-26.
    2. The elimination of mycoplasma from infected hybridomas by passaging in BALB/c mice. Carroll et al. J Immunol Methods. 1988 Apr 6;108(1-2):189-93.
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    Parameter Specification
    Appearance Red liquid
    pH 7.4 ± 0.1
    Osmolality 275-360 mOsm/L
    Endotoxin NMT< 2EU/mL
    Mycoplasma Negative
    Suitability  Suitable for mammalian cell culture
    Indicator NA
    Sterility Tested Sterile filtered using 0.22 µm filter
    Form Liquid
    Shipping Condition Dry Ice
    [/fusion_table][/fusion_builder_column][/fusion_builder_row][/fusion_builder_container]1 ml, 10 ml Read more





Mycoplasma Detection and Elimination reagents PurMa biologics certainly offers the best PCR detection kit for Mycoplasma. We also manufacture specific LB-Agar growing as well as detection of egg fried structures. The latter is as well as detection method for mycoplasma. Moreover, we manufacture mycoplasma removal and that does certainly not harm skin.

Without any shred of doubt, PurMa Biologics is the pioneering company in mycoplasma detection and elimination reagents.

Using the most sophisticated PCR detection kit, you could not only detect the mycoplasma in your cell culture system, but also determine what exact mycoplasma strain is the cause of contamination.

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