ResourceLife Sciences
A Phenol-Free DNA Extraction Method
2 May 2012In this application note by Molecular Devices, a DNA extraction method is evaluated as an alternative to a conventional phenol/chloroform extraction for pretreatment of biopharmaceutical samples. A number of protein samples were extracted using this procedure and then tested in the Threshold Total DNA Assay. This extraction procedure incorporates a chaotrope, sodium iodide, an anionic detergent, sodium N-lauroyl sarcosinate, and isopropanol to co-precipitate nucleic acids with a polysaccharide carrier, glycogen. This precipitation allows the separation of DNA from sample components, proteins, lipids, and buffers that may interfere with the Total DNA Assay. For many proteins, this extraction technique can replace a conventional overnight digest with Proteinase K and SDS, and/or a phenol/chloroform extraction.
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ElectrophoresisElectrophoresis equipment is used to separate mixtures of protein, DNA or RNA, based on their electric charge, size and other physical characteristics, by passing them through a medium such as a polyacrylamide gel, an agarose gel or a capillary tube. Electrophoresis equipment includes horizontal and vertical gel electrophoresis chambers, isoelectric focusing systems, 2D electrophoresis and capillary electrophoresis instruments. Precast gels with a gradient can be used or gels can be hand cast. Samples are run alongside ladders or markers to identify the approximate size of a molecule. Separated proteins and nucleic acids in the gel can be stained or probed with fluorescent markers and then imaged and detected using gel documentation instruments, transilluminators, densitometers and scanners. 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