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Viability PCR Starter Kits

Kits designed to make it simple to start performing viability PCR with your choice of viability dye (PMA or PMAxx™).

Viability dye
Bacteria type
Size
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price
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200 assays
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Product Description

Viability PCR starter kits contain the materials that you need for selective detection of viable cells using either PMA or PMAxx™ viability dye and real-time PCR. The kits contain Forget-Me-Not™ EvaGreen® qPCR Master Mix, and are available with or without Enhancer for gram-negative bacteria. The user will need to supply primers to amplify DNA from their species of interest (see this blog post for primer design considerations).

Features

  • Selectively detect viable cells using qPCR
  • Works even in complex samples or mixed cultures
  • Sufficient reagents to treat 80 bacterial cultures with PMA or PMAxx™
  • Sufficient reagents for 200 PCR reactions

Kit Components

  • PMA dye or PMAxx™ dye
  • PMA Enhancer for Gram Negative Bacteria (31076/31076-X only)
  • Forget-Me-Not™ EvaGreen® qPCR Master Mix
  • ROX reference dye
  • Note: User must provide primers for vPCR

About this kit

This kit contains reagents sufficient to treat 80 cultures with PMA or PMAxx™, and perform 200 PCR reactions. There is an option to choose either PMA or PMAxx™ for the viability dye. Original PMA dye has been validated for vPCR in hundreds of publications and a wide variety of bacteria, yeast, and viruses. PMAxx™ is a next-generation vPCR dye designed by Biotium scientists to provide superior live/dead discrimination compared to PMA.

Kit options are available with of PMA Enhnacer for Gram-Negative Bacteria, a reagent that improves live/dead discrimination for gram-negative bacteria (note: Enhancer is not recommended for use with gram-positive bacteria or samples with mixed strains).

To learn more about the advantages of determining microbial or cell viability using viability PCR, visit the Viability PCR Technology Page and see the PMA/PMAxx™ Reference List and Validated Bacterial Strains List.

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

Product Attributes

Viability dye
PMA, PMAxx™
Bacteria type
All species, Gram negative bacteria
Size
200 assays
Storage Conditions
Store at -10 to -35 °C
Assay type/options
Live/dead discrimination, Viability PCR
Detection method/readout
PCR/qPCR, HRM®

Documents, Protocols, SDS and COA

References

Find a list of PMA and PMAxx™ references and a list of validated bacterial strains under Supporting Documents.

FAQs

PMA and PMAxx™ for viability PCR

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

  1. 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).
  2. Let the bleach sit on the unit for 10 minutes.
  3. Thoroughly wipe the surfaces with dH2O.
  4. Wipe the surfaces with 70% ethanol and allow to air dry.

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.

For other related FAQs, see Is illumination even across all positions in the PMA-Lite™ device? and Can I use PMA or PMAxx™ with environmental samples?

PMA is stable after dilution to 0.2 mM in water as long as it is protected from light and can be stored in the same way as the 20 mM stock solution.

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