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. Additionally, the kit contains a 2X green master mix .
Mycoplasma PCR Detection Kit
$269.99 – $465.39
Description
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
- Gradual deterioration of the cell line, which is especially significant in stable cell lines (15-35%)
- Inconsistency among the results and much higher standard deviations in your statistical analysis.
- Altering DNA, RNA, and protein synthesis.
- Diminishing amino acid and ATP levels.
- Chromosomal alterations.
- Modifying host-cell immune system.
Mycoplasma contamination can happen in various ways
- 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.)
- 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.
- 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
- 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.
- Mycoplasma detection by PCR analysis. Hopert et al. In Vitro Cell Dev Biol Anim. 1993 Oct;29A(10):819-21.
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 |
Quality Control
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Background
Penicillin/Streptomycin/ Neomycin Antibiotic Mix (100X) prevents bacterial, mold & virus, mycoplasma, as well as yeast contamination. Moreover, most of the cell lines, unless they are maintained fully aseptic and come from an aseptic facility, need a mixture of antibiotics to prevent various above-mentioned contamination. Furthermore, this antibiotic mix contains all the necessary ingredients that can successfully prevent common contamination.Ingredients
Penicillin 5000 UI/ml Streptomycin 5 mg/ml Neomycin 10 mg/ml Special Features
- Cost-effective.
- Effective against both gram-positive and gram-negative bacteria.
- Contains antibiotics and antimycotics to prevent mycoplasma, fungi, and yeast.
- Provides harmless an and healthy environment for cell line if you follow the recommended concentration.
- More importantly, it does not cause toxicity level for all the available cell lines.
- Lastly, we highly recommend performing a dose-response test at the beginning of the study. That will definitely help deciding the level at which the antibiotic mix will prevent the contamination for each cell line.
Formulation
Please find the formulation here :FormulationReferences
- Screening of drugs for rapid activity against Trypanosoma cruzi trypomastigotes in vitro. Croft SL, Walker JJ, Gutteridge WE.Trop Med Parasitol. 1988 Jun;39(2):145-8.PMID:
- In vitro and in vivo studies of streptomycin-resistant, penicillin-susceptible streptococci from patients with infective endocarditis. Enzler MJ, Rouse MS, Henry NK, Geraci JE, Wilson WR. J Infect Dis. 1987 May;155(5):954-8. doi: 10.1093/infdis/155.5.954. PMID: 364485.
Mycoplasma Removal Spray
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filter_opacity_hover="100" filter_blur_hover="0" transform_type="regular" transform_hover_element="self" transform_scale_x="1" transform_scale_y="1" transform_translate_x="0" transform_translate_y="0" transform_rotate="0" transform_skew_x="0" transform_skew_y="0" transform_scale_x_hover="1" transform_scale_y_hover="1" transform_translate_x_hover="0" transform_translate_y_hover="0" transform_rotate_hover="0" transform_skew_x_hover="0" transform_skew_y_hover="0" transform_origin="" transition_duration="300" transition_easing="ease" transition_custom_easing="" animation_type="" animation_direction="left" animation_color="" animation_speed="0.3" animation_delay="0" animation_offset="" last="true" border_position="all" first="true"][fusion_text columns="" column_min_width="" column_spacing="" rule_style="" rule_size="" rule_color="" hue="" saturation="" lightness="" alpha="" content_alignment_medium="" content_alignment_small="" content_alignment="" hide_on_mobile="small-visibility,medium-visibility,large-visibility" sticky_display="normal,sticky" class="" id="" margin_top="" margin_right="" margin_bottom="" margin_left="" fusion_font_family_text_font="" fusion_font_variant_text_font="" font_size="" line_height="" letter_spacing="" text_transform="" text_color="" animation_type="" animation_direction="left" animation_color="" animation_speed="0.3" animation_delay="0" animation_offset=""]PurMa™ Mycoplasma Removal Spray
Background
PurMa™ Mycospray is the most sophisticated Mycoplasma removal spray 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
- The lack of HEPA filter and positive pressure. Without the mentioned condition, there is certainly a flow of mycoplasma in the lab.
- Moreover, lab personnel is the second main source of mycoplasma contamination.
PurMa™ Mycoplasma Removal Spray 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
- 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.
- 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.
- 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.
- On the following day clean the mentioned surfaces with 70% alcohols to remove the residues of the dead mycoplasmas.
- 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
- Wash your gloves with PurMatm Mycospray before any cell culture work
References
- 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.
- Mycoplasma removal: simple curative methods for viral supernatants. Baronti Cet al. J Virol Methods. 2013 Feb;187(2):234-7.. Epub 2012 Oct 29.
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 Antibiotic Mix Solution (Low Fungicide 2.5 µg/ml) (100X)
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box_shadow_style="" z_index_subgroup="regular" z_index="" z_index_hover="" overflow="" background_type="single" gradient_start_color="" gradient_end_color="" gradient_start_position="0" gradient_end_position="100" gradient_type="linear" radial_direction="center center" linear_angle="180" background_color="" background_image="" background_image_id="" lazy_load="none" skip_lazy_load="" background_position="left top" background_repeat="no-repeat" background_blend_mode="none" render_logics="" sticky="off" sticky_devices="small-visibility,medium-visibility,large-visibility" sticky_offset="" absolute="off" absolute_props="" filter_type="regular" filter_hover_element="self" filter_hue="0" filter_saturation="100" filter_brightness="100" filter_contrast="100" filter_invert="0" filter_sepia="0" filter_opacity="100" filter_blur="0" filter_hue_hover="0" filter_saturation_hover="100" filter_brightness_hover="100" filter_contrast_hover="100" filter_invert_hover="0" filter_sepia_hover="0" filter_opacity_hover="100" filter_blur_hover="0" transform_type="regular" transform_hover_element="self" transform_scale_x="1" transform_scale_y="1" transform_translate_x="0" transform_translate_y="0" transform_rotate="0" transform_skew_x="0" transform_skew_y="0" transform_scale_x_hover="1" transform_scale_y_hover="1" transform_translate_x_hover="0" transform_translate_y_hover="0" transform_rotate_hover="0" transform_skew_x_hover="0" transform_skew_y_hover="0" transform_origin="" transition_duration="300" transition_easing="ease" transition_custom_easing="" animation_type="" animation_direction="left" animation_color="" animation_speed="0.3" animation_delay="0" animation_offset="" last="true" border_position="all" first="true"][fusion_text columns="" column_min_width="" column_spacing="" rule_style="" rule_size="" rule_color="" hue="" saturation="" lightness="" alpha="" content_alignment_medium="" content_alignment_small="" content_alignment="" hide_on_mobile="small-visibility,medium-visibility,large-visibility" sticky_display="normal,sticky" class="" id="" margin_top="" margin_right="" margin_bottom="" margin_left="" fusion_font_family_text_font="" fusion_font_variant_text_font="" font_size="" line_height="" letter_spacing="" text_transform="" text_color="" animation_type="" animation_direction="left" animation_color="" animation_speed="0.3" animation_delay="0" animation_offset=""]Antibiotic Mix Solution (Low Fungicide 2.5 µg/ml) (100X)
Background
PurMaTM Antibiotic Mix Solution (Low Fungicide 2.5 µg/ml) (100X) generally prevents contamination of the four main pathogens in cell culture. Those are bacteria, mold/ yeast, virus as well as mycoplasma. Moreover, when cells divide in media, it carries contamination along to the daughter cells. Additionally, low fungicide antibiotic mix suites cells show sensitivity to higher amount of Amphotericin B.Ingredients
Streptomycin 10 mg/mL Penicillin 10,000 UI/mL Amphotericin B 2.5 µg/mL Special Features
- Cost-effective.
- Effective against both gram-positive and gram-negative bacteria.
- Contains antibiotics and antimycotics to prevent mycoplasma, fungi, and yeast.
- Additionally. designed to be harmless to a healthy cell line if it is used in the recommended concentration.
- More importantly, it does not cause toxicity for all the available cell lines.
- Highly recommend performing a dose-response test at the beginning to decide the level at which determines the level in which the contamination is prevented and at the same time cultured cells and tissues are in their high-performance
Formulation
For the formulation, Click here:FormulationReferences
- Risk factors for amphotericin B-induced nephrotoxicity. Luber et al. J Antimicrob Chemother. 1999 Feb;43(2):267-71. doi: 10.1093/jac/43.2.267. PMID: 11252333.
- Screening of drugs for rapid activity against Trypanosoma cruzi trypomastigotes in vitro. Croft et al.Trop Med Parasitol. 1988 Jun;39(2):145-8.
Mycoplasma Treatment Kit (MTK) (100X)
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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 Procedurebefore you start
- Decontaminate your CO2 incubator, Hood, and benches before starting the procedure, otherwise in no time, the contamination will be back into your cultured cells.
- Use unopened media bottles and fresh FBS.
- During the following mentioned procedure and after that, wear mask at all times while you work in cell culture lab.
- Spray your gloves, bench, hood with PurMa™ Mycospray (1X) (Cat#: P5M010405) before and after work. PurMaTM Mycospray (1X) instantly eliminates/ deactivates mycoplasma.
References
- Eradication of mycoplasma contaminations. Uphoff et al. Methods Mol Biol. 2013;946:15-26.
- 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.
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 Title
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