Traditional extraction methods often rely on proteinase K, SDS and purification by columns, beads or solvent extraction. Extraneous agents are added that subsequently need to be removed – all in an effort to get a simple result – high-yield, purified nucleic acids.
Much of the problem is the inadequacy of the lysis step. Proteinase K doesn’t work at a high enough temperature to denature the proteins it is digesting. To compensate, SDS is added but is toxic to every downstream diagnostic. So it must be removed, initiating a cascade of steps – each solving what was done in the previous step.
Time is wasted.
Yields aren’t preserved.
Errors are made.
Samples are vulnerable to contamination.
Technicians are exposed to harsh chemicals.
Excess plastic waste and hazardous materials need to be thrown away.
The MicroGEM Advantage
We replace proteinase K, strong detergents, and multiple steps with single-tube, hands-free, thermophilic proteinase and buffers. By controlling temperature, we effortlessly extract DNA and RNA from samples ranging from tissue, cell culture, sperm and insect, to bacteria, plant, fungi and more.
Workflow Comparison Using Temperature-Controlled Extraction vs Conventional Extraction Methods
The thermophilic proteinase is activated with a simple temperature change. The whole process is easily automated in a single tube using standard laboratory equipment – a standard thermocycler can be programmed to carry out the extraction or use MicroGEM’s PDQeX Nucleic Acid Extractor for simplified automation.
The process is a game-changer for challenging bacterial, fungal, and plant samples. The low activity of the proteinase at mesophilic temperatures means enzymes such as lysozyme, glucuronidases, cellulases and hemicellulases can be activated to break down tough cell walls. Destroyed at 75°C, they clear the way for the proteinase to do its work.
The result? Effortless nucleic acid extractions, even from challenging samples, compatible with most downstream applications.