Diploma in Lab Practices and Electrophoresis

This detailed course introduces you to the world of professional lab practices and electrophoresis. You will explore the fact that good laboratory practice (GLP) embodies a set of principles that provide a framework within which lab studies are planned, performed, monitored, recorded, reported and archived. The three levels of categorization from the Rosch Approach will be discussed. You will be introduced to the concept of buffer capacity, a quantitative measure of the resistance to change of pH by a solution, containing a buffering agent with respect to a change in the acid or alkali concentration. You will also learn that the electrophoresis in the vertical electrophoresis system performs in a discontinuous way with a buffer in the upper and lower tank, whilst the electrophoresis in the horizontal gel system performs continuously. You will discover how the calibration curve from native and denaturation gels are used to determine the molecular weight of the protein. Demonstrations, problems, examples and assessment questions in this instructor-led video-based course, are designed to provide you with a comprehensive foundation, which will enable you to take serious steps into this career.

The course begins by introducing you to the definition of GLP and the different levels of categorization within it. Then, the material safety data sheet (MSDS) will be discussed, showing you how it contains information about the potential hazards of a material. You will explore the buffer solution, which is used during experiments as a means of keeping the pH at a nearly constant value in a wide variety of chemical applications. You will discover the fact that at a fixed centrifugal force and liquid viscosity, the sedimentation rate of a particle is proportional to its size and the difference between the particle density and the density of the solution. You will also discover that electrophoresis refers to an electrokinetic process that separates charged particles in a fluid using a field of electrical charge and the reasons why the electrophoresis technique is not good for use in the separation and analysis of complex biological samples.

Next, you will learn that the complex biological samples are efficiently resolved in 2-D gel electrophoresis. This involves the combination of charge and molecular weight to provide much greater separation in comparison to the use of the individual property. You will gain insight into the fact that when conducting the agarose gel electrophoresis experiment on the native page, the sample that is prepared in the loading dye does not contain detergent or denaturing agent and as a result, the sample runs based on charge. You will then see that in the horizontal polyacrylamide gel electrophoresis, the complex biological sample is resolved according to its charge and then moved to the counter charge electrode. Study calibration curves from native and denaturation gels before moving onto liquid-handling systems, secondary structures in RNA, polyacrylamide gel electrophoresis, and stacking gel polymerization. This course will be of interest to biotechnology students, or those professionals in experimental biotechnology.