e-Myco, e-Myco plus and e-Myco VALiD

PCR Detection Kits

Detection of Mycoplasma in Cell Culture

Nasty bugs are difficult to control

Mycoplasma is a genus of bacteria which lack a cell wall. Without a cell wall, they are unaffected by penicillin or other antibiotics that target cell wall synthesis. Physically very small (< 1 micron), Mycoplasma are difficult to detect with a conventional microscope or neutralize with common antibiotics. As a result these robust bugs are often found in research laboratories as contaminants in cell culture. Reports indicate that 1/3 or more of all cell cultures are contaminated. Unfortunately, such contamination may induce cellular changes, including chromosome aberrations, changes in metabolism and cell growth. Moreover, severe Mycoplasma infections may destroy a cell line. Testing for mycoplasma is an essential quality control tool to assure accurate research and reliable biotech products.

To stop contamination before it happens, try our MycoClean Mycoplasma Prevention Spray.

PCR Process

PCR detection is the frontline for effective control

e-Myco, eMyco plus, and eMyco VALiD Mycoplasma PCR Detection Kits greatly simplify testing and detection of Mycoplasma contamination in cell cultures. With PCR testing, reliable results are obtained within a few hours, since the presence of contaminant Mycoplasmas can be easily and sensitively detected by verifying the amplified DNA fragments after gel electrophoresis. These Mycoplasma PCR Detection Kits have been shown to be a highly sensitive, specific and rapid method for the detection of Mycoplasmas contamination in cell cultures. These kits can identify the five most common cell culture-infecting species of Mycoplasma all the way down to 10 CFU/ml (e-Myco VALiD) and 20 CFU/ml (e-Myco and e-Myco plus). They also contain an internal control (primer set and template DNA) to identify false negatives caused by faulty PCR conditions and 8-methoxypsoralen (8-MOP) to prevent cross-contamination by PCR products from earlier experiments (8-MOP is known to intercalate into double-stranded nucleic acids and form a covalent inter-strand crosslink after UV activation). Each tube contains all the components for PCR (lyophilized) except for template: DNA Polymerase, dNTPs, PCR Buffer, primers set, 8-MOP, and internal control. So, you can just add your templates and perform the PCR reaction.

eMyco VALiD for the most demanding needs

e-Myco VALiD Mycoplasma PCR Detection Kit adds a measure of security when determining cell culture contamination issues. The breadth of detection is dramatically widened to include over 70 species derived from nearly 20 different host animals. And because eMyco VALiD is twice as sensitive than other eMyco kits, nary a mycoplasma can escape detection. You can get results in 3 hours, and like other eMyco kits, includes an internal control to verify a PCR run as well as positive control DNA. 8-methoxypsoralen (8-MOP) is used to permanently destroy the template activity from contaminating DNA (i.e., PCR products from earlier tests). 8-MOP is known to intercalate into double-stranded nucleic acids and form a covalent interstrand-crosslink after photo-activation by incident light at wavelength 320 – 400 nm.

Product Comparison

PRODUCT	PCR or qPCR	LYOPHILIZED PREMIX	8-MOP TO PREVENT CROSS-CONTAMINATION	# OF SPECIES DETECTED	CONTROLS	SENSITIVITY
E-Myco	PCR			49	INTERNAL	20 CFU/ml
E-Myco Plus	PCR			209	INTERNAL & EXTERNAL	20 CFU/ml
E-Myco VALiD	PCR			70	INTERNAL	10 CFU/ml

* This kit is covered by patents owned by Abbott Molecular Inc. (US Pat. No. 5,851,767 and its foreign counterparts). For research use only.