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

• PurMa^TM 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

• PurMa^TM 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.
• PurMa^TM Mycoplasma PCR Detection Kit detects the following species:  A. Laidlawii, M. Arginine, M. Boris, M. Fermentans M. Hominis, M. Hyorhinis and M. Orale
• PurMa^TM 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.