Support

Quick links
Frequently asked questions - categories
Ask a question
Contact Us
+1

PMA Real-Time PCR Bacterial Viability Kit – Salmonella enterica (invA)

This kit contains primers for amplification of Salmonella enterica invA gene, with reagents sufficient to treat 80 bacterial cultures with PMA or PMAxx™, and perform 200 PCR reactions.

Certificates of Analysis (COA)

Request COA
Viability dye

PMA, PMAxx™

Viability dye
Size
Catalog #
price
Qty
200 assays
Clear selection
Add to wishlist
ADD TO CART
(Please select quantity above )

Wishlist updated! View wishlist

Shipping, Handling & Tax (CA only) not included in price. Learn more

Tax is assessed in the state of California. All fees will be calculated during checkout based on your shipping information and are subject to change.

SPECIAL - Order 5 or more and receive 10% off.

Request a Bulk Quote

Product Description

PMA-PCR kits are designed for selective detection of viable bacteria from a specific strain using PMA or PMAxx™ dye and real-time PCR. The kits contain PMA or PMAxx™ dye, Forget-Me-Not™ qPCR Master Mix, and PCR primers for detection of selected strains of bacteria that are of widespread interest to food safety, public health, and/or antibacterial research.

This kit contains primers for amplification of Salmonella enterica invA gene, with reagents sufficient to treat 80 bacterial cultures with PMA or PMAxx™, and perform 200 PCR reactions. There is an option to choose either PMA or PMAxx™ for the viability dye (see below for more information about these dyes). The number of samples that can be treated with PMA or PMAxx™ using the kit may vary depending on sample type. See the product protocol under the downloads tab and references for more information.

Kit contents:

Strain-Specific Bacterial Viability PCR Kits

Bacteria StrainGene nameKit with PMA (catalog #)Kit with PMAxx™ (catalog #)
Salmonella entericainvA3103331033-X
Mycobacterium tuberculosisgroEL231034N/A
Staphylococcus aureusnuc31035N/A
MRSAmecA31036N/A
E. coli O157:H7Z32763103731037-X
E. coliuidA3105031050-X
Listeria monocytogeneshly3105131051-X
Legionella pneumophilamip31053N/A

PMA is a high affinity photoreactive DNA binding dye developed by Biotium. The dye is weakly fluorescent by itself but becomes highly fluorescent upon binding to nucleic acids. It preferentially binds to dsDNA with high affinity. Upon photolysis, the photoreactive azido group on the dye is converted to a highly reactive nitrene radical, which readily reacts with any hydrocarbon moiety at the binding site to form a stable covalent nitrogen-carbon bond, thus resulting in permanent DNA modification. The dye is cell membrane-impermeable and thus can be used to selectively modify DNA from dead cells with compromised membrane integrity, while leaving DNA from viable cells intact. PMA inhibits PCR amplification of modified DNA templates by a combination of removal of modified DNA during purification and inhibition of template amplification by DNA polymerases (see Reference 1 under the references tab). Consequently the dye is useful in the selective detection of viable pathogenic cells by quantitative real-time PCR.

PMAxx™ is a new and improved version of PMA designed by Biotium scientists to be a superior alternative to PMA. 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.

Forget-Me-Not™ qPCR Master Mix is a hot-start EvaGreen® dye-based master mix for use in real time PCR applications and DNA melt curve analysis. Forget-Me-Not™ master mix contains a low concentration of blue dye which allows you to see at a glance whether you forgot to add master mix to any of your tubes, so you can catch pipetting mistakes and avoid wasting time, reagents, and your precious DNA samples. It is formulated for qPCR using a fast cycling protocol, but also can be used for qPCR using regular cycling protocols. EvaGreen® dye is a unique DNA-binding dye with features ideal for both qPCR and melt curve analysis. EvaGreen® dye binds to dsDNA via a novel “release-on-demand” mechanism, which permits the use of a relatively high dye concentration in qPCR without PCR inhibition. Forget-Me-Not™ Master Mix contains Cheetah™ Taq, Biotium’s fast-activating chemically-modified hot-start Taq polymerase, which is particularly suitable for fast PCR cycling protocols.

Salmonella enterica is a gram-negative bacteria that causes the food-borne illness salmonellosis. PMA-based viability PCR for S. enterica has been reported using the primers provided in the kit (see Reference 2 under the references tab). In addition, these primers have been validated at Biotium for real-time qPCR using Forget-Me-Not™ Master Mix (see product protocol under downloads for details).

Also see our other PMA-PCR kits for detection of E. coli, E. coli strain 0157:H7, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), Listeria monocytogenes, Legionella pneumophila and Mycobacterium tuburculosis. Don’t see your favorite strain? Let us know at techsupport@biotium.com.

Materials from Biotium are sold for research use only.

References

1. Nocker A, et al. Comparison of propidium monoazide with ethidium monoazide for differentiation of live vs. dead bacteria by selective removal of DNA from dead cells. J. Microbiol. Meth. 67(2), 310-320 (2006).

2. Nocker A, et al. Molecular monitoring of disinfection efficacy using propidium monoazide in combination with quantitative PCR. J. Microbiol. Meth. 70(2), 252-260 (2007).

3. Fittipaldi M, et al. Progress in understanding preferential detection of live cells using viability dyes in combination with DNA amplification. J. Microbiol. Meth. 91(2), 276-289 (2012).

Find a full reference list under the Downloads section on this page

 

You may also like…

Support & Faq

PMA and PMAxx™ for viability PCR

Viability PCR originally used EMA (ethidium monoazide) to inactivate dead cell DNA. Biotium developed PMA (propidium monoazide) in collaboration with investigators at Montana State University (Nocker et al. 2006). PMA is more selective for dead cells than EMA, and became widely used for selective detection of viable microbes and viruses. PMAxx™ is Biotium’s newest viability PCR dye, designed to be more effective than PMA at eliminating PCR amplification of dead cell DNA. Therefore it provides the best discrimination between live and dead bacteria.

Learn more about PMA and PMAxx™ for viability PCR.

Category: PMA and PMAxx™ for viability PCR

← FAQs

Biotium offers the PMA-Lite™ LED Photolysis Device for light-induced cross-linking of PMAxx™ and PMA to dsDNA. The PMA-Lite™ is designed for samples in microcentrifuge tubes. We also offer the Glo-Plate™ Blue LED Illuminator for photoactivation of larger tubes, 96-well plates, or other plate formats.

Commercial halogen lamps (>600 W) for home use have been employed for photoactivating PMA in some publications, though results have not been consistent due to inevitable variation in the set-up configurations. If you decide to use a halogen lamp, we recommend that you lay tubes on a block of ice set 20 cm from the light source, on a rocking platform to ensure continuous mixing. Set the lamp so that the light source is pointing directly downward onto the samples (up to 45° downward slant is OK). Expose samples to light for 5-15 min.

Category: PMA and PMAxx™ for viability PCR

← FAQs

The LEDs in PMA-Lite have brightness of 600-800 millicandela (mcd). There are three LEDs next to each tube (one bottom, two side) and the wavelength is 465-475 nm.

Category: PMA and PMAxx™ for viability PCR

← FAQs

← FAQs

Ask a question

Shipping/Shelf-life

Shipping & Handling Disclaimer: Shipping and handling methods are calculated based on item temperature storage recommendation. Large, multi-item orders may be assessed additional fees. Shipping to Puerto Rico may be assessed additional fees. A Biotium customer service representative will contact you if additional fees are required. For expedited shipping, please request in the Order Notes section of the checkout page. Please note that products with recommended storage at 4°C or -20°C may ship at ambient temperature. This will not affect product performance. When you receive the product, place it under the recommended storage conditions.