DNA isolation is central to all types of biological experimentation, in that the molecule’s ability to provide defining criteria of any organism is indispensable. The first steps in the multi-isolation process involve extracting tissue and then isolating lysing cells; these ensure access to nuclear molecules, and with proper treatment, prevent the action of any enzyme that might destroy the molecules of interest. As lysis can be accomplished via a multitude of methods, here we dissect the better (more efficient) option between two that are commonly invoked: alkaline lysis and high pressure homogenization.
Alkaline lysis is largely preferred for extracting plasmid DNA from bacteria. A user would first start by growing and harvesting bacteria containing the plasmid of interest, and would follow by pelleting the cells and re-suspending them in a Tris/EDTA (or other such) solution. Lysis follows when a buffer containing NaOH and detergent is added, and the process concludes with the addition of potassium acetate to reduce the mixture’s alkalinity. (1) This method can be used for a range of sample sizes, but can be be a more costly path due to the high value of some reagents.
In contrast to that of alkaline lysis, the cell types that can be run through a high pressure homogenizer are many. From mammalian cells that only have a flexible membrane protecting intracellular contents to the bacteria and plants that also have tough cell walls, the customization option of a high pressure homogenizer allows it to accommodate each. Samples are lysed after being forced through a narrow space while being exposed to pressure, shear, and other mechanical forces. Well-suited for scaling from laboratory testing to commercial production, this method can be used for any sample size.
Ultimately, the decision to use alkaline lysis or high pressure homogenization will come down to the downstream application and other potential uses. For example, while alkaline lysis does a terrific job at lysing a single cell type well, high pressure homogenization caters to an array of cell types. So someone interested in lysing more than just bacteria may be safer with a high pressure homogenizer. Either way, it is important to conduct further research on each option to ensure a well-informed decision.
BEE International: The Homogenizer Advantage
Use of high quality cell lysis equipment will provide numerous benefits for both production laboratories and consumers alike. And there are plenty of companies on the market to select your equipment from. However, the lysate can be of higher quality and more even consistency when run through top-shelf equipment, most frequently in the form of a high pressure homogenizer. BEE International Technology is trusted by researchers and lab managers around the world. We deliver an array of key benefits, such as production of nano/micro emulsions and dispersions and lipids and suspensions; these can be used for applications such as injectables, targeted drug delivery, inhalants, time release, anesthetics, and importantly, vaccinations.
In addition, we have extensive experience in the challenges that our customers face as they transition from concept, through to R&D, clinical trials, all-important FDA approval and finally, to manufacturing. Learn more by contacting us today.
For more information about cell lysis methods and factors you should consider when choose a method, download our FREE eBook: