Blog

 

Nucleic Acid Isolation

Our understanding of genetic material has substantially increased since Friederich Miescher first extracted DNA in 1869. He discovered that a material exists within cells that precipitates out of acidic solution and dissolves into alkaline solution. He called it nuclein because it seemed to be located within the nucleus. It took until 1953 for the structure of DNA to be elucidated. It was during this time that procedures to isolate DNA began to emerge. Later, during the 1960's and 70's scientists were furiously untangling the cellular environment, and the discovery of RNA with its various forms and functions further refined DNA purification procedures. It was no longer enough to simply separate DNA from protein and salt impurities; it became necessary to remove contaminating RNA as well. Concurrently, scientists became interested in purifying messenger RNA (mRNA). Soon it became essential to purify not only DNA (genomic or plasmid), but also RNA in its various forms.

Read More
 

Differential Centrifugation in bio separation

The force of gravity will cause sedimentation of particles from a heterogeneous mixture;larger and denser particles sedimentfaster than the smaller and less dense particles. This phenomenon is useful for separating heterogeneous solutions into independent components, and for the isolation and enrichment of target molecules, cells, and cell organelles. Centrifugation accelerates the separation process by introducing centripetal forces many times greater than gravity. The precipitated particles form a pellet at the bottom of the tube during centrifugation. The rate of sedimentation is dependent on the size and density of the particles, so centrifugation can be used to isolate target particles simply by controlling centrifugal force or the rate of centrifugation. The rate of centrifugation is reported as angular velocity by revolutions per minute (rpm) or as acceleration(g). RPM is dependent on the radius of the rotor in the centrifuge.

Read More
 

Magnetic DNA purification

Nucleic acid separation can be fickle. DNA is fragile, and RNA even more so. Many commercial kits are designed to streamline the process, but they may not result in high yield or high purity DNA or RNA every time. Every laboratory is different; working habits vary, and experimental goals are not identical. It is tempting to rely on a single kit for routine isolation of genetic material because it is familiar, your lab may have published previous work with an established protocol, or you might not have the luxury or freedom to try something new. However, if you are experiencing consistently low RNA or DNA yield or purity then you may be able to justify taking time to gain a deeper understanding of the process, familiarizing yourself with the tools available, and possibly reworking your strategy.

Read More
 

Making process validation easier with a modern biomagnetic separation rack

Biomagnetic separation is a versatile and widely used tool in both industrial and small laboratory settings. It is used for the isolation of target drug molecules in the pharmaceutical realm, for the enrichment of enzymes in industry, and for in-vitro diagnostics in medicine. It is especially useful in the small-scale research environment for inexpensive target cell enrichment, protein isolation, or nucleic acid capture. In the early days of biomagnetic separation it was thought that the process was only reliable for small volumes. However, the development of modern biomagnetic separation racks has made it possible to scale up the process to large volumes and to enable process validation and consistency between batches.

Read More
 

Enzymes in Industry

 

Enzymes play a surprisingly important role in modern industry, and are essential to the production of more commercial products than one would initially consider. Enzymes are proteins that speed up reactions and improve yield by increasing available precursors for downstream reactions. Perhaps the most obvious use for enzymes in industryisthe production of cheese, bread, and alcohol. In these traditional applications the enzymes are part of microbial machinery such as bacteria or yeast. Over time scientists have been able to isolate specific enzymes and to understand their catalytic functions well enough to incorporate them with or without their microbial hosts into a wide variety of somewhat surprising situations. For example, enzymes are usedinthe production of textiles, detergents, biofuels, and pharmaceutical products. Large quantities of desired enzymes are required for these applications, and they need to be available in the purest form possible. The purity of enzymes in industry is particular important for pharmaceutical applications where the products as well as the process are susceptible to review and control by regulatory associations. Batches of enzymes in industry undergo regular process validation to ensure batch-to-batch consistency.

Read More
 

Filtration systems for Production and R&D

The ability to obtain an enriched population of small molecules, cells, proteins, nucleic acids, or contaminant-free solutions is important for all applications: from small laboratory research up to the large-scale production of pharmaceutical products. Filtration systems are available in all shapes, sizes, and materials for diverse situations. The need to meet regulatory guidelines for purity and consistency of pharmaceutical productsdemands a well-designed enrichment plan and filtration system.

Read More
 

Magnetic properties of nanoparticles

Magnetic nanoparticles are gaining traction as a useful tool in a variety of applications including drug delivery, therapeutic treatment, contrast agents for MRI imaging, bioseparation, and in-vitro diagnostics.  These nanometer-sized particles are useful because they are superparamagnetic, which is a direct result of their tiny size—only a few nanometers—a fraction of the width of a human hair (nanoparticles are approximately 1/1,000 thinner than human hair). Superparamagnetic nanoparticles are not magnetic when located in a zero magnetic field, but they quickly become magnetized when an external magnetic field is applied. When returned to a zero magnetic field they quickly revert to a non-magnetized state.

Read More
 

Properties of magnetic nanoparticles: synthesis, protection, functionalization, and application

Part 2: Structure/Protection, Functionalization, and Application

The first part of this series provided a general overview of the most common synthesis methods for generating superparamagnetic nanoparticles of only a few nanometers in size. This second part touches on the procedures necessary to protect and functionalize these nanoparticles to extend their usefulness across a great number of applications.
Read More
 

Magnetic immunoprecipitation (ip) input into western blot analysis

Immunoprecipitation

Immunoprecipitation (ip) is a technique for capturing specific proteins via antibody-antigen affinity from a complex solution. A co-ip, instead of identifying individual proteins, is designed to identify protein complexes. The phrase “pulling down” protein is commonly used to explain the process, but this idea is somewhat dated now that magnetic nanoparticles have begun to replace traditional centrifuge-based methods. The protein capture efficiency can be measured by ip input ito SDS page and western blot analysis.

Read More
 

Properties of magnetic nanoparticles: synthesis, protection, functionalization, and application

 

Part I: Synthesis


Magnetic nanoparticles have risen in popularity in medical and biotechnology fields over the past decade. These tiny nanometer-sized particles are superparamagnetic, which means they can be magnetized by an externally applied magnetic field and quickly returned to a non-magnetic state once the field is removed. They are easy to manipulate, making them perfect for biomagnetic separation processes and a variety of other applications. There are many options to consider when choosing a magnetic nanoparticle for an experiment or therapeutic goal. A general understanding of the synthesis, protection, functionalization, and application of magnetic nanoparticles is a good place to start. In the first part of this series we focus on the synthesis of magnetic nanoparticles.

Read More

Leave a comment