The e-Myco™ VALiD-Q Mycoplasma qPCR Detection Kit is designed for the highly sensitive, efficient detection of Mycoplasma through real-time qPCR. The kit contains a set of primers and a probe specifically developed for detecting the highly conserved Mycoplasma 16S-rRNA coding region.
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.
With the e-Myco™ VALiD-Q Mycoplasma qPCR Detection Kit, over 70 species of Mycoplasma can be detected including M. pneumoniae, M. argnini, M. hyorhinis, M. fermentans, M. orale and A. laidlawii. The detection process consists of first extracting DNA from the specimen and then amplifying the extracted DNA using a 5' nuclease fluorescent probe and a specific primers pair. This assay amplifies two type specific regions: Mycoplasma (FAM) and IPC (HEX). Internal controls are included in the Mycoplasma Detection solution for monitoring the extraction process and identifying PCR inhibition. Positive control DNA is included as well. Detection can be achieved in 90 minutes with a sensitivity up to 10CFU/ml.
Two protocols are provided with this kit (see below). The extraction step in protocol B involves Intron's G-spin Total DNA Extraction Kit for optimized DNA purification from a variety of specimens including animal cell cultures, blood, animal tissue, rodent tail, fixed tissue, hair and Gram-negative bacteria.