Viability PCR With PMA and PMAxx™ dyes
- Used to selectively modify exposed DNA from dead cells with compromised membrane integrity
- Makes the dye useful for the detection of viable pathogenic cells
- PMA yields better live/dead discrimination than ethidium monoazide bromide (EMA)
All-In-One Kits
Cells can be treated with PMA or PMAxx™ prior to any quantitative PCR reaction, which is ideal for users that already have their an established PCR assay for their strain of interest. However for maximal convenience, we also offer bacterial strain-specific viability PCR kits for several popular bacterial strains. These kits are designed for the selective detection of viable bacteria from a specific strain using a viability dye (PMA or PMAxx™) and real-time PCR. The kits contain either PMAxx™ or PMA viability dye, our exceptionally sensitive Forget-Me-Not™ qPCR Master Mix, and a set of validated PCR primers for detection of selected strains of bacteria that are of widespread interest to food safety, public health, and/or antibacterial research. Kits for Gram-negative strains also include PMA Enhancer. Kits contain reagents sufficient to treat 80 bacterial cultures with PMA or PMAxx™, and perform 200 PCR reactions.
Propidium monoazide (PMA) is cell membrane-impermeable. It therefore can be used to selectively modify exposed DNA from dead cells with compromised membrane integrity, while leaving DNA from viable cells intact. This feature makes the dye useful for the detection of viable pathogenic cells in the presence of dead cells by quantitative PCR. PMA modification inhibits PCR amplification of DNA from dead cells, allowing selective qPCR amplification of unmodified DNA from viable cells. PMA is less membrane permeable than ethidium monoazide bromide (EMA), the traditionally used reagent for viability PCR, and therefore yields better live/dead discrimination.
PMA Enhancer for Gram Negative Bacteria was designed to improve PMA-mediated discrimination between live and dead gram-negative bacteria. PMA Enhancer is provided as a 5X solution, and is added to a sample before the addition of PMA or PMAxx™. When a sequence from a gram-negative bacteria is amplified by PCR, samples pre-treated with Enhancer show a decrease in the signal from dead cells, with no change in the signal from live cells. Thus, PMA plus Enhancer is the optimal way to perform viability PCR on gram-negative bacteria. Note: PMA Enhancer is not intended for use when gram-positive bacteria are to be detected. PMA Enhancer may adversely affect the amplification of live cell DNA from gram-positive bacteria.
PMA-Lite™ LED Photolysis Device is specifically designed for photoactivation of propidium monoazide (PMA), PMAxx™, and EMA. It can also function as a general photolysis device to provide continuous illumination to 1.5-2 mL-sized vials in a controlled manner. The device can hold up to 18 vials. Multiple LED lights are positioned to provide even and maximal illumination from both the sides and bottom to all vials. An internal fan is used to reduce the heat generated from the lights to ensure near room temperature photolysis. Each PMA-Lite™ device includes a free vial of PMAxx™. PMA-Lite LED Photolysis device is CE marked.
• Holds up to 18 microcentrifuge tubes
• Bright, long-lasting LEDs with 465-475 nm emission
• Designed for efficient photoactivation of PMA, PMAxx™ and similar dyes
• Internal fan to ensure temperature below 37°C
• Four timer settings
Glo-Plate™ Blue LED Illuminator. A multi-functional blue LED light box that can be used for photolysis of PMAxx™ or PMA in viability PCR. It is also an excellent way to develop the staining of gels stained with the visible blue DNA gel stain DNAzure®.
• Flat illumination surface fits microplates
• Bright, long-lasting LEDs with 465-475 nm emission
• Designed for efficient photoactivation of PMA, PMAxx™ and similar dyes
• Surface stays cool during light exposure
• Four timer settings
Ordering information: PMA dye is available in solid form or preformulated as a 20 mM solution in dH2O.
Caution: Store at ‒20°C and protect from light at all times.
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