MBI evolution Mycoplasma Real time PCR Detection Kit
Mycoplasmas are small, round or filamentous prokaryotic organisms which are a frequent contaminant of cell cultures. Mycoplasma depend on their hosts for many nutrients due to their limited biosynthetic capabilities. Up to 30~85% of cell cultures may be contaminated with mycoplasmas, the main contaminants being the species M.orale, A.laidlawii, M.arginini and M.hyorhinis. Although these mycoplasmas do not usually kill contaminated cells, they are difficult to detect and can cause a variety of effects on cultured cells, including changes in metabolism growth, viability and morphology, there by altering the phenotypic properties of the host cells.
Many methods are available for detection of mycoplasma, including isolation in broth/agar culture, direct or indirect fluorescence staining, immunostaining, direct or indirect PCR. Among those methods, direct PCR is the highly sensitive, specific and convenient method when the primer design is optimized. MBI evolution Mycoplasma Real time PCR Detection Kit is composed of a set of primers and probe that are specific for the highly conserved mycoplasma16S-rRNA coding region including M.pneumoniae, M.argnini, M.hyorhinis, M.fermentans, M.orale and A.laidlawii.
The kit is designed to detect the presence of mycoplasma that might contaminate biological materials such as cultured cells. Also, the kit can detect mycoplasma within 90minutes sensitively up to 10CFU/ml and includes internal control for verifying a qPCR run as well as positive control DNA
- The real-time qPCR(quantitative polymerase chain reaction) DNA amplification technology shows high sensitivity and specificity for direct detection of pathogen (antigen). iNtRON developed a novel platform technique about primer design called CLP™ (complementary locking primer) technology which provides flexibility in Tm (melting temperature) of primer design for optimization of reaction condition, and maximizes PCR specificity and sensitivity through the control of non-specific priming.
- The kit contains all reagents required for real-time polymerase chain reaction: nucleotides, primer and probes, reaction buffer, polymerase and controls. After rehydration of the components the PCR Master mix is easily prepared by just adding the Inhibition Control DNA and Taq DNA polymerase to the Primer & Probes.
- The PCR is initiated with a 5 minute high-temperature step to melt all nucleic acids and to activate the polymerase. In a successful PCR, the presence of mycoplasmas will be indicated by a signal in the FAM channel.