e-Myco, e-Myco plus, and e-Myco VALiD PCR Detection Kits

Detection of Mycoplasma in Cell Culture

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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 , 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 (e-Myco VALiD) and (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 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 () 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




E-Myco PCR
20 CFU/ml
E-Myco Plus PCR
20 CFU/ml
10 CFU/ml

Additional Information

Genomic DNA was isolated from M.fermentans-infected K562 using a DNA extraction minikit. The isolated gDNA was serially diluted for PCR of mycoplasma detection. These results show that it can be applied to mycoplasma detection with small quantities, such as 3.25 pg of gDNA.

Lane M, 100bp DNA Marker; lane IC, Internal control, lane 1, 50ng; lane 2, 25ng; lane 3, 12.5ng; lane 4, 6.3ng; lane 5, 3.2ng; lane 6, 1.6ng; lane 7, 800pg; lane 8, 400pg; lane 9, 200pg; lane 10, 100pg; lane 11, 50pg; lane 12, 25 pg; lane 13, 12.5 pg; lane 14, 6.3 pg; lane 15, 3.25 pg

*Similar results were seen with K562 cell (M. fermentans-infected ) using as little as 12 cells.

Lane M, SiZer™-100 DNA Marker; Lane 1, 1×106 cfu/ml of gDNA; Lane 2, 1×105 cfu/ml of gDNA; Lane 3, 1×104 cfu/ml of gDNA; Lane 4, 1×103 cfu/ml of gDNA; Lane 5, 1×102 cfu/ml of gDNA; Lane 6, 10 cfu/ml of gDNA; Lane 7, 1 cfu/ml of gDNA; Lane N, No template control.

To determine the minimal required concentration of Mycoplasma, M. fermentans was selected as a model Mycoplasma species. M. fermentans was serially diluted for each PCR reaction. The results indicates that the detection limit with this kit is 20 cfu/ml.

Elimination of cross-contamination using 8-MOP activation

by UV irradiation of 1st PCR Product

Panel A – Lane M, SiZer™-1000 plus DNA Marker; lane 1, Internal control; lane 2, 50pg; lane 3, 10pg; lane 4, 2pg; lane 5, 0.4pg

Panel B – lane M, SiZer™-1000 plus DNA Marker ; lane 1, 5, PCR product (1 μl) used from lane 1 of panel A; lane 2, 6, PCR product (1 μl) used from lane 2 of panel A; lane 3, 7 , PCR product (1 μl) used from lane 3 of panel A; lane 4, 8, PCR product (1 μl) used from lane 4 of panel A.

Storage and ease of use are two often overlooked factors when working with reagents for molecular biology. When all other things are equal, it’s nice to know that your reagents can be just as effective on day 360 as on day 1. And better yet, kits designed for minimum pipeting is just a smarter approach. iNtRON has applied an elegant solution to these challenges. A proprietary freeze-dried technique utilized in most of their molecular biology kits stabilizes the entire master mix at the bottom of 0.2 ml PCR tubes. Then each strip of tubes is individually vacuum sealed to guarantee that no water infiltration can spoil the master mix over time. Simply add sample (and/or primers) and your experiments are off to the races. Eliminating preparation of master mixes and other components frees up time and reduces errors. Whether it’s PCR kits, diagnostic kits, or other enzyme kits, iNtRON products have incredibly long shelf lives for even the most finicky enzymes. We want you to be confident every time you purchase a PCR premix from us!

Contents PCR kits – 48 or 96 x 20µl rxns in 8-strip lypholized tubes


Part Numbers 2523348, 25233, 2523448, 25234, 2523148, 25231
Master Mix Yes, just add template
Detection Method Agarose Gel (Standard PCR Kits)
Target Size ~260 bp
Number of Species Detected eMyco = 49
eMyco plus = 209
eMyco VALiD = 70
Detection Limit 20 CFU/ml (e-Myco & e-Myco plus)

10 CFU/ml (e-Myco VALiD)

Internal Control Yes, ~160 bp
Cross Contamination Control Yes, 8-methoxypsoralen (8-MOP) for PCR kits only.
Total Protocol Time <3 hours
Storage 1 year at -20°C

* 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.