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Venor®GeM qOneStep is a mycoplasma qPCR detection kit which includes all reagents required for the qPCR reaction. Primer, nucleotides, polymerase and the internal amplification control are provided ready-to-use in a lyophilized reaction mix. Rehydration buffer and lyophilized Positive Control DNA are also provided in the kit. The assay can be performed with any type of real-time PCR cycler able to detect the fluorescence dyes FAM™ and HEX™. The protocol provided is recommended for the fast and reliable screening of cell culture supernatants. The detection procedure can be performed within 3 hours.
Mycoplasmas are specifically detected by amplifying a highly conserved rRNA operon, or more precisely, the 16S rRNA coding region in the mycoplasma genome. The mycoplasma-specific amplification is detected at 520 nm (FAM™ channel). The qOneStep PCR reaction mix includes primer sets and FAM™ labeled probes which allow the specific detection of all Mollicute species so far described as contaminants of cell cultures and media components. Eukaryotic DNA is not amplified by this primer/probe system. False negative results due to PCR inhibitors or improper DNA extraction are detected by the internal amplification control which is already included in qOneStep reaction mix. The amplification of the internal amplification control is detected at 610 nm (HEX™ channel).
TaqMan® based qPCR Assay with FAM™ and HEX™ labeled probes
Applicable in research and development for direct testing of cell cultures and biologicals. Intended for research use only. Not recommended for clinical diagnostics or testing of human samples.
Primer sets, nucleotides, internal amplification control and polymerase at optimized concentrations, lyophilized and pre-cast in one tube
Rehydration Buffer
Lyophilized Positive Control DNA
PCR grade water
qOneStep PCR reaction mixes are prepared in aliquots of 25 reactions each.
cycler-based, real-time PCR
PCR reaction tubes
qPCR cycler
Pipetting equipment
Microcentrifuge
Venor®GeM qOneStep components can be stored at +2 to +8 °C for at least 12 months. After rehydration, reagents must be stored at ≤ 18 °C.
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