Cell culture contamination control: Detection

Venor®GeM qOneStep

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.

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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).

Type of PCRTaqMan® based qPCR Assay with FAM™ and HEX™ labeled probes

Scope of ApplicationApplicable 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.

Kit ComponentsPrimer sets, nucleotides, internal amplification control and polymerase at optimized concentrations, lyophilized and pre-cast in one tubeRehydration BufferLyophilized Positive Control DNAPCR grade water

Package SizesqOneStep PCR reaction mixes are prepared in aliquots of 25 reactions each. Cat. No. 11-91025 25 tests Cat. No. 11-91100 100 tests Cat. No. 11-91250 250 testsResult evaluationcycler-based, real-time PCR

Required ConsumablesPCR reaction tubes

Required lab devicesqPCR cyclerPipetting equipmentMicrocentrifuge

Shelf Life and StorageVenor®GeM qOneStep components can be stored at2 to8 °C for at least 12 months. After rehydration, reagents must be stored at ≤ 18 °C.EP 2.6.7 complianceNo

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Manual and Instructions for use

quick visual instructions

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Technical notes

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