What is a polyacrylamide gel?
polyacrylamide gel is a solution commonly used in electrophoresis, the process of separating molecules or particles of different sizes by passing gel and using electric current. There are various recipes for polyacrylamide gel, but usually contains acrylamide, water, buffer, persulpate ammonia (APS) and tetramethylemendiamine (TEMED). This gel acts as a matrix that separates compounds based on their charge and size; The more acrylamide in the gel solution, the better the separation of smaller molecules. This gel is usually used to separate protein and DNA.
In the electrophoresis, the electrical fields of charged particles that move through a gel that act as a sieve, causing the particles of different sizes to separate. The gel absorbs any heat produced from the electric current. In this procedure, polyacrylamide gel is commonly used, but can also be used agarosis, other chemicals that create a similar gel.
CAN gels must be made of different concentrations, with a lot of acrylamideFrom about 6% to 15%. When mixing the gel, acrylamide is non -bodylamerized, which means it remains as individual molecules. The addition of other chemicals that promote binding, such as Timed, causes molecules to connect long chains together - "polyacrylamide.
The main advantage of polyacrylamide gel is that the number of links and hardness can be controlled on the basis of the initial amount of acrylamide and added. The main factor of acrylamide concentration is the size of the analyzed molecule. The smaller the compounds are separate, the more acrylamide is needed; Very small particles are separated by the highest concentration, 15 percent.
acrylamide works by checking the amount of friction in the gel; Gel hardness determines the amount of fraction. Compounds that are large move the gel more slowly, because due to their size is naturally greater friction or resistance. Smaller compounds move rmore, because it is less friction. To avoid moving small compounds from the gel, more acrylamide is added to create more friction.
DNA polyacrylamide gel is used to separate DNA chains with different lengths. The pieces of bottom will be separated by only one nucleotide, compounds that make up each spring. To know which pieces refer to specific sizes, a standard that contains fragments of known size is commonly performed with samples. DNA pieces are then compared to the standard and the string size is determined.
A similar procedure is used to determine the size of proteins, most often in the blood. Blood contains two main types of protein: globulin, large size protein and serum albumin, very small, negatively charged protein. Separation using polyacrylamide gel is USED to determine the amount of each type.