Alkaline Lysis vs. High Pressure Homogenization: Which is Better?

Posted by David Shechter on Dec 7, 2016 12:30:00 PM

Alkaline Lysis.jpgIn the world of biotechnology – more specifically, the field of molecular biology – the process of lysing cells is an important (and often complicated) task. There are two common methods of accomplishing the goals of lysing cells – alkaline lysis and high pressure homogenization. Continue reading to learn about both methods, as well as which method would work best for your industry’s needs.

Alkaline lysis – also known as alkaline extraction – is the process of isolating plasmid DNA from bacteria. Alkaline lysis is popular due to its ease, speed, and reliability. While methods can vary from lab to lab, the general process of alkaline lysis is as follows:

Spinning Down the Cells

The first step in alkaline lysis involves spinning down the cells in order to concentrate cellular material. This is often done in a centrifuge containing the sample as well as a supernatant.

Discarding the Supernatant and Resuspending the Cells

After the sample is centrifuged, the supernatant is discarded and the desired cells are resuspended in buffer.

Lysing the Cells

After resuspension, the cells are then able to be lysed. The lysing agent – often sodium hydroxide – works to disrupt and lyse the cells.

While alkaline lysis is relatively simple, this method is best suited for smaller scale production and does not offer much flexibility for alternate applications. High Pressure Homogenizers, like the ones offered at BEE International, offer the same speed, ease, and reliability of alkaline lysis, with the following additional benefits:

  • Ability to control pressure in order to lyse a variety of cell types, including more challenging cells such as yeast or fungi
  • Accessibility of intracellular proteins
  • Built-in process flexibility
  • High yields in less time
  • Various sample size processing
  • Rupturing cells without damaging the intracellular materials
  • Better results in fewer passes
  • Scalable results, for both small and large operations

Our equipment is suitable for a variety of applications, from small scale lab use, all the way to larger applications, such as pilot plants and 24/7 manufacturing facilities. Our high pressure homogenizers include the Nano DeBEE, Micro DeBEE, and Mini DeBEE for laboratory use, as well as DeBEE 2000, DeBEE 3000, and DeBEE 4000, suitable for pilot plant and industrial use.

For more information about any of our high pressure homogenization equipment, as well as information on how our products can best suit your needs, please contact us. In addition, our free eBook, “7 Key Factors to Consider When Choosing a Cell Lysis Method” can be downloaded for free here.

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