PMAxx™ Dye, 20 mM in H₂O

Viability PCR dye, a new and improved version of the popular viability dye propidium monoazide (PMA).

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100 uL

1 mL

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PMAxxâ¢ Dye, 20 mM in H<sub>2</sub>O - Image 2

Viability PCR dyes like PMAxx™ or PMA are membrane-impermeant, which makes them dead cell specific. Once inside of a dead cell, they bind to DNA. Exposure to intense visible light renders the dyes reactive, and causes them to covalently attach to the DNA. This DNA modification prevents amplification in subsequent PCR reactions.

PMAxxâ¢ Dye, 20 mM in H<sub>2</sub>O - Image 3

The basic v-PCR workflow involves the following 5 steps: dye addition, incubation, light exposure, DNA extraction, and qPCR.

PMAxxâ¢ Dye, 20 mM in H<sub>2</sub>O - Image 4

Live or dead bacteria treated with either PMA or PMAxx™ prior to qPCR. Dead bacteria treated with either PMA or PMAxx™ show delays in amplification compared to either live cells, or untreated dead cells. Bacteria treated with PMAxx™ show a larger delay than those treated with PMA. Therefore PMAxx™ is better able to differentiate live vs dead bacteria by viability PCR.

PMAxxâ¢ Dye, 20 mM in H<sub>2</sub>O - Image 5

E. coli were killed with mild heat treatment (56°C for 3 hrs) and treated with PMAxx™ or PMAxx™ + Enhancer, followed by light exposure using PMA-Lite™, DNA purification, and qPCR with Fast EvaGreen® qPCR Master Mix. dCt values were calculated by subtracting the Ct without dye from the Ct with dye. Only dead cells treated with PMAxx™ + Enhancer showed a large dCt, indicating that the dye successfully inhibited PCR of dead cell DNA.

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Product Description

PMAxx™ is a photoreactive DNA-binding dye for viability PCR. It was designed by Biotium scientists to be a superior alternative to the popular viability dye propidium monoazide (PMA).

Selectively detect viable cells using qPCR
Better live/dead discrimination of bacteria than PMA
Dead cell specific dye, binds to DNA
Covalently attaches to DNA after photoactivation
20 mM in water, 100 uL volume
λ_Abs = 464 nm (before photolysis); λ_Abs/λ_Em= ~510/~610 nm (with DNA/RNA, after photolysis)

To learn more about the advantages of determining microbial or cell viability using viability PCR, visit the Viability PCR Technology Page.

About PMAxx™

PMAxx™ dye is a DNA modifier used for viability PCR, invented by scientists at Biotium. PMAxx™ is a new and improved version of our popular viability dye PMA (propidium monoazide). Like PMA, PMAxx™ is a photo-reactive dye that binds to DNA with high affinity. Upon photolysis with visible light, PMAxx™ dye becomes covalently attached to DNA. This modified DNA cannot be amplified by PCR. PMAxx™ dye is designed to be cell membrane-impermeant. Thus, in a population of live and dead cells, only dead cells are susceptible to DNA modification due to compromised cell membranes. This unique feature makes PMAxx™ highly useful in selective detection of live bacteria by qPCR.

PMAxx™ was designed by Biotium scientists to be a superior alternative to PMA, and is compatible with all existing PMA protocols, but with superior activity. While PMA is generally effective at differentiating between live and dead bacteria by qPCR, it does not completely eliminate PCR products from dead cell DNA. This could potentially give false positive results. Biotium’s new dye PMAxx™ is much more effective at eliminating PCR amplification of dead cell DNA, and therefore provides the best discrimination between live and dead bacteria.

PMAxx technology is covered by granted and/or pending US and international patents.

Product Attributes

Size

100 uL, 1 mL

Viability dye

PMAxx™

Storage Conditions

Store at -10 to -35 °C

Excitation/Emission

Abs = 464 nm (before photolysis), ~510/~610 nm (with DNA/RNA, after photolysis)

Assay type/options

Live/dead discrimination, Viability PCR

Detection method/readout

PCR/qPCR

Documents, Protocols, SDS and COA

Documents

PMA & PMAxx™ References

PMA & PMAxx™ Validated Bacterial Strains

Bacteria Stain Flyer

PMAxx™ & PMA Flyer

Cell Viability and Cell Death Brochure

Protocols

PI-40069

SDS

SDS 40069

Datasheet

Datasheet Download

COA

Request the Certificate of Analysis

References

Download list of curated PMA and PMAxx™ References and a list of PMA and PMAxx™ Validated Bacterial Strains.

Citations

Download list of curated PMA and PMAxx™ References and a list of PMA and PMAxx™ Validated Bacterial Strains.

FAQs

PMA and PMAxx™ for viability PCR

What is the procedure for decontaminating the PMA-Lite™ 2.0 LED Photolysis Device?

Note: Do not remove the cover or introduce liquids to the interior of the PMA-Lite.

Thoroughly wipe all exposed PMA-Lite surfaces and the inner rims of the tube holes with 10% bleach in water (household bleach diluted at a ratio of 1 part bleach to 9 parts water).
Let the bleach sit on the unit for 10 minutes.
Thoroughly wipe the surfaces with dH2O.
Wipe the surfaces with 70% ethanol and allow to air dry.

What is the wavelength and brightness (luminosity) of the LED lights in the PMA-Lite?

The LEDs in the PMA-Lite™ and PMA-Lite™ 2.0 have a wavelength that is 465-475 nm and a brightness of approximately 600-800 millicandela (mcd). These are nominal values provided for reference use only, individual LED wavelength and brightness are not a calibrated specifications for the device.

There are three LEDs in each well (one bottom, two side) that provide illumination around each sample tube for efficient photoactivation.

The illumination in each well on the PMA-Lite far exceeds what is required for photocrosslinking of the viability dyes EMA, PMA, or PMAxx™ to nucleic acids. Therefore, any variability in brightness of the PMA-Lite LEDs should not significantly affect the v-PCR results. If performance verification is required, we recommend doing a functional PMA-PCR assay to verify that PMA-treated samples photoactivated in the device give qPCR results within an acceptable range. Mixing the samples during photoactivation and using longer illumination times may be necessary if the samples are complex and not fully transparent to light.