PMA qPCR Live Bacteria Detection Kit (without Primers)

PMA qPCR Live Bacteria Detection Kit facilitates bacteria activity detection. This kit provides PMA dye and SYBR Green qPCR premix. The optimal using amount and sample amount vary according to  sample type. PMA is a DNA binding dye with high affinity, especially with double-stranded DNA. This dye has weak fluorescence it self but exhibits brighter fluorescence after binding with nucleic acid. PMA is impermeable to cell membrane, therefore is able to selectively modify DNA in dead cells with impaired membrane. PMA-modified DNA PMA-modified DNA undergoes photolysis upon exposure to blue light (~464 nm), converting the photoreactive azide group on PMA into a highly reactive azene radical. This radical reacts with any nearby hydrocarbon moiety near the DNA binding site to form a stable covalent nitrogen-carbon bond, resulting in permanent DNA modification. This modification process renders DNA insoluble and causes it to be lost along with cellular debris during subsequent genomic DNA extraction. When exposed to strong light, the remaining unbound PMA will react with water and decompose into hydroxylamine compounds lacking cross-linking activity, unable to covalently bind with DNA. Based on this property, we combined PMA with qPCR technology and developed a novel detection method: PMA-qPCR for live bacteria screening. Currently, this method has been validated in various bacteria strains and yeast, fungi, virus, and parasites. Complex samples such as feces or soil, the treatment onmay require optimization of dilution, dye concentration, light treatment period. For diluted samples such as tested water, they may require filtration or concentration before dye treatment.