Buffer Solutions Prepared with Ultrasonication

Lysis buffers can be efficiently and quick prepared using ultrasonic tissue homogenizers. Since ultrasonicators are a dream tool for mixing, dissolving and dispersing, they allow for a reliable and user-friendly preparation of lysis buffers.

Ultrasonic Preparation of Lysis Buffers

Ultrasonic tissue homogenizers are a common tool for cell disruption, lysis and extraction, therefore available in most laboratories. A secondary application for probe-type ultrasonicators is the preparation of lysis buffers.

Lysis buffer is a solution used to break open (lyse) cells and release their contents for downstream analysis. The specific composition of lysis buffer can vary depending on the type of cells being lysed and the downstream application. However, a typical lysis buffer contains components such as detergents, salts, and protease inhibitors. Buffering salts (e.g. Tris-HCl) and ionic salts (e.g. NaCl) are commonly used to regulate the pH and osmolarity of the lysate.
To make a lysis buffer, the components are mixed together in the appropriate concentrations and pH. The mixture is usually stirred or shaken until the components are dissolved and the solution is homogeneous.
Ultrasonic homogenization is a method that can be used to help make a good lysis buffer by improving the mixing and dissolution of the components. In this method, ultrasound waves – normally in the range of 20 to 30 kHz – are applied to the mixture, which creates cavitation bubbles that collapse and generate intense local energy, leading to mechanical shear and mixing of the components.
The ultrasonic homogenization process can help to break apart any clumps or aggregates of the components, and ensure that the solution is uniformly mixed. Consequently, such improved lysis buffer can lead to more efficient cell disruption and higher yields of extracted material. Additionally, ultrasonic homogenization can reduce the time required to make a lysis buffer compared to traditional stirring or shaking methods.
Overall, ultrasonic homogenization is a powerful tool for improving the quality and efficiency of lysis buffer preparation.

Protocol for Ultrasonic Lysis Buffer Preparation

This is a general protocol for making a lysis buffer using an probe-type ultrasonicator:
 
Materials:

• Detergent(s) of choice (e.g. Triton X-100, SDS, CHAPS)
• Salt(s) of choice (e.g. NaCl, KCl)
• Protease inhibitors (e.g. PMSF, aprotinin, leupeptin)
• Ultrasonicator
• Stirrer
• Deionized water
• pH meter or pH strips

 
 
Step-by-step Instructions:

1. Determine the composition of the lysis buffer based on the cells or tissue you will be lysing, and the downstream applications you will be using the lysate for. Prepare the stock solutions of detergents, salts, and protease inhibitors at the desired concentrations.
2. Add the stock solutions of detergents, salts, and protease inhibitors to a beaker or flask.
3. Add deionized water to bring the volume up to the desired amount of buffer.
4. Mix the components using a stirrer in order to get a pre-mix solution.
5. Measure the pH of the buffer using a pH meter or pH strips, and adjust the pH as necessary with small amounts of acid or base.
6. Use the probe-type ultrasonicator to homogenize the pre-mix solution so that a highly homogenous solution is obtained. Therefore, sonicate the solution for a few seconds until the solution is thoroughly mixed and any clumps or aggregates are broken down. The duration and power of sonication can be optimized based on the specific lysis buffer and ultrasonicator used.
7. After ultrasonication, re-measure the pH of the buffer and adjust as necessary.
8. Filter the buffer through a 0.2 micron filter to remove any debris or aggregates that may have formed during sonication.
9. The lysis buffer is now ready for use.

 
Note: The exact composition and pH of the lysis buffer can vary depending on the cells or tissue being lysed and the downstream applications. The above protocol is a general guideline and may need to be optimized for specific applications.
 
Ultrasonic homogenizers are commonly used for cell disruption and extraction of intracellular molecules. Therefore many lysis applications use probe-type sonication not only for the preparation of the lysis buffer, but also for the mechanical cell disruption und extraction.
This makes ultrasonic tissue homogenizers a versatile tool for laboratories and explains the wide use of ultrasonicators in microbiology, biotechnology and life science.

Ultrasonic Lab Homogenizers

Hielscher Ultrasonics manufactures and supplies ultrasonic homogenizers and probes (sonotrodes) at various power ratings and sample volumes. This allows us to offer you the most suitable ultrasonic disruptor for your sample preparation. We focus on highest quality, outstanding user-comfort and reliable sonication results. All digital ultrasonicators feature smart software, programming and pre-setting of sonication protocols, browser remote control, temperature control, sample illumination as well as automatic data recording on a built-in SD-card.

Design, Manufacturing and Consulting – Quality Made in Germany

Hielscher ultrasonicators are well-known for their highest quality and design standards. Robustness and easy operation allow the smooth integration of our ultrasonicators into industrial facilities. Rough conditions and demanding environments are easily handled by Hielscher ultrasonicators.

Hielscher Ultrasonics is an ISO certified company and put special emphasis on high-performance ultrasonicators featuring state-of-the-art technology and user-friendliness. Of course, Hielscher ultrasonicators are CE compliant and meet the requirements of UL, CSA and RoHs.

The table below gives you an overview of our various lab ultrasonicators: