What Is Cell Lysis & How Can You Make the Process More Efficient?

Posted by Deb Shechter on Nov 2, 2016 12:30:00 PM

cell-lysis.jpgCell lysis is the rupture of the cell membrane resulting in the release of cell contents, and the subsequent death of the cell. Cell lysis can occur naturally, for example, through a viral infection or osmosis. In research laboratories and in industry, cell lysis is often used to break open the cell and study its contents.

Cell lysis can be achieved through mechanical or chemical methods. Mechanical cell lysis techniques include high shear mixing, traditional homogenization, sonication, grinding, freeze/thaw cycles, and high pressure homogenization. Chemical cell lysis techniques include osmotic lysis, and the use of detergents, chelating agents, or chaotropic agents.

The success of a cell lysing process depends on several factors. Firstly, the process should be flexible, as different cells require different cell lysis strategies. The process should be relatively easy to perform, and result in a high yield in a short time. Lastly, results should be consistently reproducible and scalable.

Making Cell Lysis More Efficient

The first step in efficient cell lysis is choosing the correct method for your particular sample. What works for one type of cell may not work as well (or at all) for other cell types. You should also take your resources (time, budget, skill level of laboratory staff etc.) into account when deciding on a method.

Take the freeze/thaw method, for example. This method involves freezing a cell suspension and then allowing the material to thaw at room temperature. This causes the cells to swell and ultimately break as ice crystals form during the freezing process and then contract during thawing. Multiple cycles are often necessary, making the process quite time-consuming. While this method is suitable for lysing bacterial cells and algae, it is not very effective for harder plant materials, which may instead require a stronger mechanical force like a mortar and pestle or a tissue grinder.

DeBEE High Pressure Homogenizers: Versatile and Effective

BEE International’s unique, modular technology allows you to gently rupture cells without damaging the valuable intracellular materials. You are able to control the pressure, allowing for rupture of a variety of cell types. No harsh chemicals are introduced into the process, and all results are 100% scalable to manufacturing.

For more information on cell disruption, download our free eBook: 7 Key Factors to Consider When Choosing a Cell Lysis Method or contact us today!

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Cell Homogenization: Types of Cells & The Best Way to Lyse Them

Posted by Deb Shechter on Oct 28, 2016 12:30:00 PM

cell homogenization Cell lysis (or cell homogenization) is the rupture of the cell membrane resulting in the release of cell contents, and the subsequent death of the cell. Cell lysis takes place in biotech, pharmaceutical, food, cosmetic, and chemical laboratories every day. Research scientists and laboratory technicians disrupt the cell’s structure in order to extract organelles, proteins, DNA, or mRNA. However, many common cell lysis methods are incomplete, leading to wasted time, excess costs, high risks, and they can even undermine the entire research effort.

A wide variety of cells require rupture such as E. coli, yeast, mammalian tissue, bacteria, algae, fungi and insect cells. The primary difference between plant and animal cells is that plant cells have a cell wall and a cell membrane, while animal cells only have a cell membrane.

Adaptable Cell Homogenization with DeBEE High Pressure Homogenizers

Different types of cells and biological materials require different cell disruption methods. Some only require gentle processing, while others need harsher methods. Sonicators, for example, are unsuitable for lysing certain plant cells, as cell walls are very resistant to sonication. Organic solvents like alcohols, ether or chloroform can disrupt cells by permeating the cell walls and membranes. These solvents are often used, in combination with shearing forces, to lyse plant cells.

BEE International offers a flexible approach to cell homogenization, as the process is adjustable to handle a wide variety of cells. Our modular technology allows you the ability to control the pressure in order to rupture different cell types including more challenging cells such as yeast or fungi. Cells are ruptured without damaging the valuable intracellular materials, and without the use of solvents and chemicals.

DeBEE High Pressure Homogenizers use forces like shear, cavitation and impact to disrupt cells. Different cells can be ruptured using the most efficient process for the highest recovery rates.

Advantages of Cell Homogenization with DeBEE High Pressure Homogenizers:

  • Accessibility of intracellular proteins
  • Built-in process flexibility
  • Ease-of-use and low learning curve
  • High yields in less time
  • Reproducible and scalable results
  • Various sample size processing
  • Ability to disrupt all cell types
  • Better results in fewer passes

For a quote on any one of our homogenizer models, contact us today and let us show you the advantage of BEE High Pressure Homogenizers for your application. For more information on which cell lysis method is best for you, download our FREE eBook: “7 Key Factors to Consider When Choosing a Cell Lysis Method”:

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Cell Rupture: Detergents vs Traditional Physical Methods

Posted by Tal Shechter on Sep 26, 2016 12:30:00 PM

cell ruptureDetergents for Cell Rupture

Detergents (or surfactants) are used in cell lysis solutions because they disrupt the distinct interface between hydrophobic and hydrophilic systems. They help to solubilize membrane proteins and lipids, thereby causing the cell to lyse and release its contents.

Detergents are comprised of a polar hydrophilic head group and a nonpolar hydrophobic tail. They are categorized by the nature of the head group as either ionic, nonionic or zwitterionic.

Nonionic and zwitterionic detergents are generally milder and less denaturing than ionic detergents. Examples include CHAPS (zwitterionic) and Triton (nonionic). Ionic detergents are considered to be harsh detergents. They are strong solubilizing agents and tend to denature proteins. Examples include soap or alcohol ethoxysulfates (anionic detergents), and quaternary ammonium compounds (cationic detergents).

Physical Methods of Cell Rupture

There are several methods that are commonly used to physically lyse cells, including:

  1. Mechanical disruption: Using various equipment to cut, chop, grind and crush the sample.
  2. Sonication: Using pulsed, high frequency sound waves to lyse cells. This process can be direct (a probe is inserted into the sample) or indirect (the energy is transmitted through a bath of water into the sample vessels).
  3. Freeze-Thaw method: This technique involves freezing a cell suspension and then thawing the material at room temperature. This causes cells to swell and ultimately break as ice crystals form during the freezing process and then contract during thawing. The process is repeated as necessary.
  4. Homogenization: The sample is forced through a very narrow nozzle. The higher the amount of energy applied during the homogenization process, the more efficient the cell lysis.

Which one to choose?

Detergent-based lysis is a popular method for cell rupture. It is fairly easy to do and does not require any special equipment. However, detergent cell lysis is often too mild, and needs to be done in conjunction with a physical method like grinding or homogenization. Also, bear in mind that harsh detergents can often damage or destroy the contents of the cell if used incorrectly.

Our high pressure homogenizing technology at BEE International allows you to gently rupture cells without damaging the valuable intracellular materials. No harsh chemicals are introduced into the process, and all results are 100% scalable to manufacturing.

Still unsure? Download our free eBook: 7 Key Factors to Consider When Choosing a Cell Lysis Method or contact us here.

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Cell Lysis Techniques: Why Homogenization Excels

Posted by David Shechter on Sep 7, 2016 11:30:00 AM

cell lysisCell lysis (or cell disruption) is the rupture of the cell membrane resulting in the release of cell contents, and the subsequent death of the cell. The fluid containing the contents of lysed cells is called a lysate.

Cell lysis is used in laboratories to separate the intracellular contents, e.g. in DNA or RNA extraction. Cell lysis is also a key component in protein purification. Protein purification involves a series of processes to isolate one or a few proteins from the rest of the lysate.

Cell lysis disturbs the carefully controlled cellular environment, allowing for the possibility of damage to the intracellular contents. Proteases (enzymes that break down proteins) are also released upon cell lysis, so the process should be kept quick, the lysate must be kept cool, or protease inhibitors should be added to the lysis buffer.

Mechanical cell lysis techniques include high shear mixing, traditional homogenization, sonication, grinding, and high pressure homogenization. Chemical cell lysis techniques include osmotic lysis, the use of detergents, chelating agents, or chaotropic agents.

Key Factors for Successful Cell Lysis Techniques:

  • Intracellular proteins should be accessible for extraction and solubilization.
  • The process should be flexible, as different cells require different cell lysis strategies.
  • The process should be relatively easy to perform.
  • The ideal technique results in a high yield in a short time.
  • Results should be consistently reproducible and scalable.

High Pressure Homogenization: A Superior Cell Lysis Technique

DeBEE high pressure homogenizers are able to gently lyse a wide variety of cells. This is because our proprietary technology allows you to control the process and vary the pressure according to cell type. Cells are ruptured, but without disturbing the valuable intracellular material within.

Our modular equipment allows for a flexible process and independent control of pressure, flow, cavitation, impact, shear, and process duration. At the heart of our proprietary technology is our BEE homogenizing cell. This supports powerful, repeated use of ALL mixing forces (turbulence, cavitation, shear and impact). Our equipment is easy to use, produces better results in fewer passes, and results are always scalable.

BEE: “Best Emulsifying Equipment”

At BEE International, we are passionate about product innovation and are committed to providing the best possible results for your product or application. We are experienced in every aspect of high energy mixing; and all of our equipment reflect our dedication to advancing the technology of homogenization. 

Contact us to learn more about our cell lysis solutions, and check out our free eBook: 7 Key Factors to Consider When Choosing a Cell Lysis Method.

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Cell Lysis Sonication: Is It Better Than Homogenization?

Posted by David Shechter on Sep 1, 2016 12:30:00 PM

cell lysisWhat is Cell Lysis Sonication?

Sonication uses pulsed, high frequency sound waves to lyse cells. There are two kinds of commonly used sonication techniques:

  1. Direct sonication: A titanium probe is inserted into the sample vessel. Ultrasonic energy is transmitted directly from the probe into the sample, allowing for rapid processing. This unit is called a sonicator (click here for a quick demonstration).
  2. Indirect sonication: Can be performed with an ultrasonic bath or a cup horn sonicator. This method does not require a probe that comes into direct contact with the sample; thus lowering the possibility of cross contamination. The ultrasonic energy is transmitted through a bath of water into the sample vessels.

Both direct and indirect sonication are able to lyse cells through cavitation. This phenomenon occurs when a rapid pressure change causes “liquid-free zones” or cavities in a liquid. As these cavities collapse, shock waves are generated throughout the liquid, which causes the cells to rupture.

Limitations of Sonication

One disadvantage of cell lysis sonication is the generation of heat. To prevent excessive heat, the ultrasonic pulses are applied in short bursts to a sample that is immersed in an ice bath. This process can be time consuming, and sonication is only a viable option for small volumes.

Another drawback to sonication is that it does not lyse all cells equally. For example, cell walls are far more resistant to cell lysis sonication than cell membranes, making plant cells more difficult to rupture than animal cells.

Cell Lysis with DeBEE High Pressure Homogenizers

De BEE high pressure homogenizers are uniquely suited to gently rupturing a wide variety of cells. Our proprietary technology allows you to control the process and vary the pressure according to the cell type. Cells are ruptured, but without disturbing the valuable intracellular material within.

Our high pressure homogenizers also make use of cavitation (just like cell lysis sonication) but we ALSO use the forces of shear and impact. Cell lysis is gentle but effective, allowing for better results in fewer passes. Best of all, our results are consistently scalable up to manufacturing volumes. We have several different high pressure homogenizer products for your laboratory, pilot plant or production facility.

Contact us today for a quote or to discuss your cell lysis needs.

Download our free eBook “7 Key Factors to Consider When Choosing a Cell Lysis Method” to learn more about how to select the right cell lysis equipment for your application.

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Homogenizers for Optimal Cell Lysis & Particle Size Reduction

Posted by Deb Shechter on Aug 26, 2016 12:30:00 PM

high pressure homogenizerHigh pressure homogenization is a mechanical process that works to reduce particle size or to lyse cells. Essentially, a liquid is forced at high pressure through a very narrow nozzle. The higher the amount of energy applied during the homogenization process, the smaller the particle size or the more complete the cell lysis.

Below, we’ll discuss the forces used to apply this energy; as well as the differences that set DeBEE high pressure homogenizers apart from the competition.

Our unique BEE International technology begins right at the product intake, which can be laminar and gentle, or turbulent for premixing. As the liquid enters the homogenizer nozzle, there is a sharp drop in pressure, causing cavitation. Cavitation occurs when a rapid pressure change causes “liquid-free zones” or cavities in a liquid. As these cavities collapse, shock waves are generated throughout the liquid, which causes the particles to break apart.

The liquid continues on into an absorption chamber which contains alternating small and large orifices, which creates turbulence and causes fluid-on-fluid impact and shear. As a result of the application of these forces (cavitation, turbulence, impact and shear), the liquid that now flows out of the homogenizer has a much smaller particle size than before.

The DeBEE Difference

The above process describes a basic high pressure homogenizer set up, but our DeBEE high pressure homogenizers offer several additional benefits:

  • Process Intensity and Duration: Our high pressure homogenizers can be configured with a reverse flow setup, where the high velocity liquid jet is made to reverse on itself and to exit near the nozzle, thus causing even more impact and fluid-on-fluid shear. The duration of the process can be controlled by the length and internal configuration of the absorption chamber. Process intensity is adjustable from 2000 - 45000 psi / 150 - 3100 bar.
  • Constant Pressure:  A constant processing pressure leads to a more uniform particle size. Our unique High Pressure Pumping System uses several hydraulically driven intensifier pumps for a constant, uninterrupted processing pressure. This pumping system also eliminates the possibility of contamination. Read more about it here.
  • Modular Technology: BEE International’s proprietary technology allows independent control of pressure, flow, cavitation, impact, shear, and process duration. This allows you to optimize cell lysis or particle size reduction and obtain the best possible results for your product.

Contact us today to request a quote or a demonstration and experience the DeBEE difference! For more information about particle size reduction and how to achieve efficient and consistent results, download our FREE eBook:

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Chemical Methods of Cell Disruption

Posted by David Shechter on Aug 24, 2016 12:30:00 PM

cell disruptionCell disruption, or cell lysis, is achieved when the cell wall or membrane is ruptured, releasing the contents of the cell. Cell disruption is the first step in many biotechnology applications. This sensitive process needs to be controlled as much as possible; and choosing the correct cell lysis method is essential for preserving the desired intracellular contents.

Cell disruption can be achieved in several ways, including mechanical, enzymatic or chemical lysis. Read on to learn more about chemical methods of cell disruption:

The very simplest form of chemical cell lysis is osmotic lysis. Here, cells are suspended in a hypotonic extracellular environment (often a dilute sucrose solution). This causes the cells to take on water, swell and subsequently burst. Osmotic lysis does not occur in plant cells due to their sturdy cell walls. Organic solvents like alcohols, ether or chloroform can disrupt cells by permeating the cell walls and membranes. These solvents are often used (in combination with shearing forces) with plant cells.

EDTA (ethylenediaminetetraacetic acid) is a chelating agent which can be used to disrupt gram negative microorganisms, since it chelates the cations, leaving holes in the cell walls.

Surfactants (commonly called detergents) disrupt the distinct interface between hydrophobic and hydrophilic systems.  Detergents are used in cell lysis buffers and they help to solubilize membrane proteins and lipids thereby causing the cell to lyse and release its contents. Widely used detergents include Triton and sodium dodecyl sulfate (SDS).

Chaotropic agents, such as urea and guanidine, are also used for cell lysis. They are capable of bringing some hydrophobic compounds into aqueous solutions. They do this by disrupting the structure of water and making it a less hydrophilic environment, and weakening the hydrophobic interactions among solute molecules.

Disadvantages of Chemical Methods of Cell Disruption:

A major drawback in using chemical methods of cell disruption in manufacturing is the cost. Using small amounts of chemicals and enzymes in the R&D laboratory is acceptable; but the cost of using the large volumes required for large-scale production is often not feasible.

Harsh chemicals and detergents can often damage or destroy the contents of the cell if used incorrectly. Lastly, using large volumes of potentially hazardous chemicals creates significant health and safety risks.

The Benefits of Mechanical Cell Disruption with High Pressure Homogenization:

Our high pressure homogenizing technology at BEE International allows you to gently rupture cells without damaging the valuable intracellular materials. You are able to control the pressure, allowing for rupture of a variety of cell types. No harsh chemicals are introduced into the process, and all results are 100% scalable to manufacturing.

For more information on cell disruption, download our FREE eBook: 7 Key Factors to Consider When Choosing a Cell Lysis Method or contact us today!

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Alkaline Lysis vs. High Pressure Homogenization: Which Lyses More Efficiently?

Posted by David Shechter on Jul 26, 2016 12:30:00 PM

high pressure homogenizationDNA 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:

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Dounce Homogenizer: Better than High Pressure Homogenization?

Posted by Tal Shechter on Jul 13, 2016 12:30:00 PM

Dounce_HomogenizerMechanical homogenization is oftentimes preferable to other methods in its ability to impart physical change without altering the chemical properties of the sample. Cell lysis, an oft-visited homogenizer application, exemplifies this concept; while chemical disruption methods risk altering the intracellular molecules of interest, mechanical disruption methods simply cause a physical break in the membrane and/or wall while leaving subcellular fractions injury-free. Dounce homogenizers and high pressure homogenizers (HPH) can both be used to mechanically lyse cells, but which is better, particularly for your intended application?

The concept of homogenization was brought into existence in the 1800’s, when scientists began altering milk’s properties to improve its taste and physical appearance. The first homogenizer was patented in 1899 by Auguste Gaulin, and since then, this simple machine has evolved significantly. (1) The Dounce homogenizer and the HPH are great examples of this model because of their similar purpose and vast procedural difference. The Dounce homogenizer has a quite simple setup, in that it is comprised of a glass mortar and pestle with a miniscule clearance between the two, which allows for a stress-induced lysis of cells and tissues. In contrast, HPH forces a sample through a narrow tube while imparting multiple mechanical forces to rupture the cells. Although each is an effective method of homogenization, they  represent a distinct evolutionary lineage from the first homogenizer.

Aside from their basic mechanisms of action, Dounce homogenizers and HPH cater to a very different crowd. Firstly, the number of steps should be taken into consideration; an ideal technique has one single step and HPH generally meets this format. HPH simply requires setting of the dials for customization and entry of the sample into the machine. While the sample may require pre- or post-homogenization treatment, the homogenization process itself has that sweet spot of easy setup and a machine completing the work. On the opposing end, Dounce homogenization is one of multiple manual steps that are required to effectively lyse a cell. For example, a scientist would first cut tissue to isolate subcellular fractions, coarsely grind the sample, and then use a Dounce homogenizer for final rupture. (2) Moreover, Dounce homogenizers can process only small samples at a single time and require significant user participation with the mortar and pestle. HPH can process samples on both a small and large scale, making it more valuable to scientists.

BEE International: High Pressure Homogenizers That Effectively Disrupt Cells

Researchers who require cell lysates, DNA, RNA, or protein extract should have access to a high pressure homogenizer. On determining which homogenizer will be the best fit, the search can begin with BEE International Technology. We are globally recognized among laboratory managers and researchers for our high quality products and excellent customer support. Cell lysis is just one of a variety of applications for our homogenizers; nano/micro emulsions, lipids, suspensions, and dispersions are also easily achievable. Additionally, the homogenizer processes can be controlled to suit any given product, which will allow for customization to the cell type. And finally, the equipment is easy to use, produces higher yield in less time, and achieves results that are reproducible and scalable.

Learn more about how to effectively lyse your cell sample by contacting us today. Also, if you are looking for more information on cell lysis methods and how to choose the right one for you, download our FREE eBook: 

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How Homogenizers Efficiently Lyse E. coli Cells

Posted by Tal Shechter on Jun 8, 2016 12:30:00 PM

Lyse E. Coli cellsFrom synthetic fuel and medicine to experimentation for DNA and evolution education, use as a cancer treatment tool and construction of bio-computers, Escherichia coli (E. coli) is critical to the success of many industries. (1) Its cells contain proteins with widespread life science applications, as well as DNA with an impressive number of genes considering its small size. Access to these precious materials requires cell lysis and preparation of the lysate, of which a variety of methods are available. Most commonly used is homogenization, which offers numerous benefits over other methods. Read on to learn about how homogenizers can efficiently lyse E. coli cells.

So how does it work its mixing magic? When a liquid E. coli suspension is inserted into a homogenizer, it is pushed through a narrow tube at high pressure. The sudden release of pressure that follows provides mechanical force, specifically turbulence and shear, which rupture the cells. Interestingly, the amount of cell breakage achieved is in direct proportion to operating pressure; this fact emphasizes the importance of using a machine that can achieve comparatively high operating pressures. The desired product, whether protein, gene, or other intracellular molecule, will dictate specifically what is extracted and how it is prepared after homogenization concludes.

As intriguing as homogenization may be sounding at this point, it is prudent to consider other mixing options, of which there are many. For example, freeze-thawing, centrifugation, chemical disruption, and sonication are all viable options. Yet compared with each of these, homogenization confers significant benefits like higher disruption rates, increased efficiency in terms of time and outcome, and enhanced ability to scale. Additionally, homogenization is a good fit for both small and large sample sizes, and its use of force instead of temperature change (like heat or freeze-thawing) preserves intracellular structures for extraction.

BEE International: Homogenizers for Effective Cell Disruption

As your laboratory prepares to disrupt E. coli cells for protein extraction, you will begin looking at homogenizers that best suit the required needs. BEE International is trusted by pharmaceutical researchers and lab managers around the world. We deliver an array of key benefits, such as production of cell lysates, nano/micro emulsions and dispersions and lipids and suspensions. Our homogenizer processes can be controlled to better suit your product; for example, pressure can be adjusted to be gentler or harsher and the results can be scaled to manufacturing. Finally, our equipment is easy to use, produces higher yield in less time, and results are reproducible and scalable.

Learn more about how BEEI can assist you in cell disruption by contacting us today. For more information about cell lysis and what you should consider when choosing the right method for you, download our FREE eBook:

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