The PCR Mycoplasma Test Kit I/RT utilizes quantitative, real-time PCR (qPCR). It shows excellent sensitivity (detects approximately 10 mycoplasmas per sample volume) and specificity and allows to obtain reliable PCR results extremely fast (in “real time”). Thus, the kit is suited particularly well for routine testing/screening for mycoplasma contamination using cell culture supernatants. It contains mycoplasma-specific probes and comes with Inhibition Control DNA as well as Positive Control DNA. Defined DNA standards for quantitative Real-Time PCR are available separately.

The kit detects a wide range of mycoplasma species and is compatible with any Real-Time PCR instrument able to detect the fluorescence dyes FAM™ and HEX™. The complete detection procedure can be performed within 3 hours.

Background

Mycoplasmas are specifically detected by amplifying a highly conserved 23S rRNA coding region in the mycoplasma genome. This mycoplasma-specific amplification is detected at 520 nm (FAM™ channel).The kit includes a lyophilized, complete Master Mix containing primer and FAM™ labeled probes which allow the specific detection of all Mollicutes 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 can be added to the PCR master mix. The amplification of the internal amplification control is detected at 560 nm (HEX™ channel).

Kit Components PCR Mycoplasma Test Kit I/RT; Variant C– Primer&Probes Mix: containing mycoplasma specific primer set; nucleotides dATP, dCTP, dGTP and dUTP; Taq DNA Polymerase; (lyophilized master mix; for 25 or 50 reactions)– Inhibition Control Spike: containing artificial DNA fragments/plasmid DNA (lyophilized; non-infectious; for at least 25 or 50 reactions)– Positive Control Spike, containing artificial DNA-fragments/plasmid DNA (lyophilized; non-infectious; for at least 25 or 50 reactions)– Rehydration/Reaction Buffer– DNA-free Water– Instruction manual

Mollicutes species detectable with PCR Mycoplasma Test Kit I/RT, Variant C| Based on positive testing | Based on sequence alignments | Based on sequence alignments ||—————————|——————————|——————————|| Mycoplasma arginini | Mycoplasma agalactiae | Mycoplasma hyopharyngis || Mycoplasma arthritidis | Mycoplasma alligatoris | Mycoplasma hyopneumoniae || Mycoplasma fermentans | Mycoplasma alvi | Mycoplasma hyosynoviae || Mycoplasma gallisepticum | Mycoplasma amphoriforme | Mycoplasma iguanae || Mycoplasma genitalium | Mycoplasma anatis | Mycoplasma imitans || Mycoplasma hominis | Mycoplasma bovis | Mycoplasma mobile || Mycoplasma hyorhinis | Mycoplasma canis | Mycoplasma ovipneumoniae || Mycoplasma orale | Mycoplasma columbinum | Mycoplasma pulmonis || Mycoplasma penetrans | Mycoplasma conjunctivae | Mycoplasma testudineum || Mycoplasma pneumoniae | Mycoplasma crocodyli | Mycoplasma testudinis || Mycoplasma salivarium | Mycoplasma flocculare | || Mycoplasma synoviae | | || Acholeplasma laidlawii | | |