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Reaction buffers for photo reactive dyes

Reaction buffers for combining with photo reactive dyes ( i.e Propidium or Ethidium Monoazide)

One of the critical steps in all viable cell detection workflow by the means of PMA or EMA photoreaction, is sample and reagent incubation on darkness. During this treatment is very important to maximize reagent diffusion trough dead cells, spores, and cysts of microbial aggregates.

On the other hand also is necessary to prevent viable cell damage, for this reason ionic strength, pH, nutrient level and aerobic/anaerobic balance needs to be under control. GenIUL introduces three different reaction buffers: Standard , Standard Plus and Anaerobic.

These reaction buffers are compatible with vPCR, Flow Cytometry and Fluorescence Microscopy procedures.

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Information

The PhAST is thermally stable with a constant and uniform light dose. It allows simultaneous photo activation of 12 samples in a simple and efficient manner.

PhAST combines high power LED with the proper optical alignment of the reaction tube to ensure the maximum efficiency in the binding of the reagent to DNA.

PhAST allows optimizing photo labeling method by programming different parameters such as light intensities and photo-activation times.

In the detection and enumeration of microorganisms in clinical samples, environmental or food control by molecular biology techniques it crucial to differentiate live cells from dead cells. The assessment and understanding of microbial reality may have large biases and alter our conclusions.

The use of reagents will turn the applicability of genetic techniques and applications in many fields, including:

Clinical Microbiology and infectious disease evolution.
Pathogen Detection and health risk assessment of environmental and food samples.
Susceptibility studies: biocides, toxic, antibiotics.
Microbial ecology.

In this new era, the resolution of molecular techniques can be improved with the living cells differentiation.

Unique Features

Bifidobacterium cells supspensions with Reaction buffer anaerobic or standard, were treated with a conventional vPCR workflow. ( 50 mM of PMA during 30 min, PhAST Blue at 100% during 15 min).
Reaction buffer anaerobic protects anaerobic cells during vPCR work flow.
Thermaly inactivated (95ºC, 30 min) Mycobacteria suspension.
Untreated with PMA or treated with 50 mM, 30 min.
with Reaction Buffer standard or Reaction Buffer plus.
PhAST Blue photoactivation 10 min, 100%.
Reaction Buffer Plus completely reduces qPCR signal
Reaction Buffer standardFor general purposes, neutral pH, standard ionic strength.
Reaction Buffer plusContains anti clumping agents, cell membrane fluidising agents and a reduced level of nutrients. Designed for stabilizing live cells but accelerating reagent diffusion trough dead cells. Compatible with Enterobacteria and Mycobacteria.
Reaction Buffer anaerobicContains anti clumping agents, minimum nutrient levels and provides a reducing environment compatible with PMA or EMA based reagents. Designed for stabilizing anaerobic live cells but accelerating reagent diffusion trough dead cells. Compatible with Bifidobacteriaceae

One of the critical steps in all viable cell detection workflow by the means of PMA or EMA photoreaction, is sample and reagent incubation on darkness. During this treatment is very important to maximize reagent diffusion trough dead cells, spores, and cysts of microbial aggregates.