PMA-PCR kits are designed for selective detection of viable bacteria from a specific strain using PMA or PMAxx™ dye and real-time PCR.
- This kit provides primers for specific amplification of the Salmonella enterica invA gene
- Sufficient reagents for treating of 80 bacterial cultures with PMA or PMAxx™
- Sufficient reagents for performing 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 and references for more information.
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. 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
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 firstname.lastname@example.org.
Strain-Specific Bacterial Viability PCR Kits
Materials from Biotium are sold for research use only.
Find a list of PMA & PMAxx references and a validated strains list under Supporting Documents.