Electrophoresis is the separation of particles caused by an electric current. Scientists use this process to examine humans and other species at the genetic level. The process separates the components of DNA to determine the presence of certain genetic genes and even the presence of disease and disease. The test's relatively low cost and extreme accuracy have allowed science to map genetic markers for various blood-borne disorders, including sickle cell anemia.
Identification Versatility
Electrophoresis is a general diagnostic test that can be used for protein and nucleic acid separation or protein separation fingerprinting. The system is commonly used in the medical field to diagnose various genetic blood disorders such as sickle cell anemia and thalassemia in a one-dimensional or two-dimensional test to isolate proteins and identify genetic markers. Electrophoresis has also been used to identify different species of fish and to determine valuable properties of soybeans and wheat by analyzing and separating constituent DNA.
Low cost of use
The cost of electrophoretic analysis is about $2.60 per gel sample analyzed, or less than 3 cents per point of data, according to estimates in Crop Science, a journal of genomics, molecular genetics and biotechnology published by Michigan State University . In terms of genetic research and analysis, this makes electrophoresis an inexpensive means of identifying genetic markers on a large scale using machines with up to 100 samples at a time.
accuracy of results
Electrophoresis is very precise. When this process is performed correctly, it can divide the proteins present inside the cell into as many as 1500 different parts. The system is also highly selective because it is able to notice differences in DNA samples, even when those samples differ by only two base pairs. This provides reliable test results that allow scientists and lab technicians to draw conclusions about the genetic makeup of the species being tested to grow tougher crops, determine the possible presence of a disease in humans, and perhaps one day be able to identify genetically Eliminate the disease horizontally.
MT4
In molecular biology, good buffer selection and preparation for different DNA isolation steps can be the difference between continuing with protein expression or opening another maxi-prep kit to start over. While the importance of buffer recipes cannot be overstated, recipes are usually just that - recipes are written with no explanation or rationale for the different components. One such buffer is glucose-tris-EDTA or GTE buffer.
What is GTE used for?
GTE is used to lyse (break open) the cells and resuspend the bacterial cell pellet before harvesting the plasmid DNA within. Lysozyme, which dissolves cell membranes, is usually added with GTE buffer. Obtaining a homogenous suspension of intact cells at this step so that subsequently added lysates can access all cells is critical for good DNA yields. GTE aims to do this while also providing a stable environment for DNA.
glucose osmolarity
Add 50 mM (millimolar) glucose to the GTE buffer to maintain osmolarity at which the concentration of solutes outside the cell approaches that inside the cell. This prevents premature lysis of cells, which can lead to lower DNA yields due to aggregation and degradation. Other components of the buffer also contribute to the osmolarity of the solution, but glucose as a non-electrolyte is a good choice because it does not interfere with the buffer properties of the solution.
pH Stable Tris
Tris is short for tris(hydroxymethyl)aminomethane, which is a very common pH buffer. For GTE buffer, salt (Tris-HCl) was added to the buffer at a concentration of 25 mM. This keeps the pH of the solution close to physiological 8.0, a desirable pH to prevent acid hydrolysis (degradation) of the plasmid DNA and unwanted side effects of other cellular components.
EDTA prevents DNA degradation
EDTA, or ethylenediaminetetraacetic acid, traps or "chelates" metal ions from solution, preventing them from participating in unwanted side reactions. In GTE buffer, EDTA was added at 10 mM. Its main purpose is to collect free zinc, magnesium, and calcium in the buffer, thereby preventing DNA degradation through certain pathways that require these metals.
some important tips
Keep the GTE buffer cold and keep it in small quantities to prevent accidental bacterial growth in it. Sugar and a controlled pH make a good growth medium. Always use purified water. Tap water pipes may contain excess metal ions, which may overwhelm EDTA's capture capacity. If you suspect interfering RNA in your sample, add RNase A to your GTE buffer at 100 µg/ml to eliminate this problem.
