e-Myco VALID Mycoplasma PCR Detection Kit

The e-Myco™ VALID Mycoplasma PCR Detection Kit consists of a set of primers specially designed to detect the highly conserved mycoplasma 16S-rRNA coding region, including M. pneumoniae, M. agninini, M. hyorhinis, M. fermentans, M. orale and A. laidlawii. The species of mycoplasma can be determined by sequencing the amplified products. These tests have high sensitivity and are suitable for release testing and in-process control. They can replace culture and indicator cell tests.

Mycoplasma is a common and serious contaminant of cell cultures. It has been shown that 30-87% of cell cultures are infected with Mycoplasma. In continuous cell cultures, contaminating Mycoplasma may grow slowly, not killing the cells outright but affecting various parameters, leading to unreliable results. Testing for Mycoplasma is an important and necessary quality control measure. 

The primers used in this Mycoplasma detection kit anneal to conserved regions of the Mycoplasma genome, allowing detection of all important species of Mycoplasma. Mycoplasma contaminants can be detected simply as amplified fragments on a DNA gel; labeled probes are not necessary.

Each e-Myco VALID Mycoplasma PCR detection tube contains all components required for PCR in a dried and stable pellet format, including i-StarTaq DNA Polymerase, dNTPs, 10x Buffer, Mycoplasma primers, 8-methoxypsoralen, and an internal control for Mycoplasma partial gene amplifications. The reaction can begin by simply adding the template DNA or samples to be tested. PCR with this primer set is based on conserved 16S rRNA and delivers detection results in only a few hours. This is far more efficient than the conventional detection methods that involve culturing samples on selective media and require at least 1 week to obtain results. 

The e-Myco tubes contain an adopted 8-methoxypsoralen to extinguish the template activity of contaminated DNA. 8-methoxypsoralen is known to intercalate into double-stranded nucleic acids and form a covalent interstrand crosslink after photo-activation by incident light at wavelengths of 320-400 nm. An internal control of this product was constructed to identify false negative results in each reaction. The internal control causes the sample primer pair to amplify the internal control and target DNA, which are differentiated by size.