Gel Electrophoresis and Molecular Sciences
Gel electrophoresis is one of the main techniques in molecular biology. The basic principle of this technique is that DNA, RNA, or protein can be separated by an electric field. In this case, the molecules are separated based on their rate of movement by the electromotive force in the gel matrix.
The primers are single-stranded oligonucleotides whose sequences are designed complementary to the end of the DNA fragment to be copied; The primary determines the beginning and end of the area to be copied. The last step is the elongation, which is the elongation of the primer into a new DNA strand by the DNA polymerase enzyme.
The temperature at this stage depends on the type of DNA polymerase used. Eventually, one PCR cycle will double the number of printed DNA or target DNA molecules, because each new synthesized thread will act as a template for the next cycle.
The rate of displacement depends on the size of the molecule concerned. Gel electrophoresis is usually performed for analytical purposes, but can also be used as a preparative technique for purifying molecules before being used in other methods such as mass spectrometry, PCR, cloning, DNA sequencing, or immuno-blotting which are further characterization methods.
The gel used is usually a crosslinked polymer (crosslinked) whose porosity can be adjusted as needed. To separate small proteins or nucleic acids (DNA, RNA, or oligonucleotides), the gel used is usually a polyacrylamide gel, made with different concentrations between acrylamide and substances that allow cross-linking, resulting in polyacrylamide networks of different sizes cavities vary. To separate larger nucleic acids (greater than several hundred bases), the gel used is agarose (from seaweed extract) which has been purified.
In the electrophoresis process, a molecular sample is placed into a well in a gel placed in a buffer solution, and electricity is supplied to it. The sample molecules will move in the gel matrix in the direction of one of the electric poles according to their charge. In the case of nucleic acids, the direction of movement is towards the positive electrode, due to the natural negative charge on the sugar-phosphate frame it has
. To keep the rate of transfer of nucleic acids really only by size (ie length), substances such as sodium hydroxide or formamide are used to keep nucleic acids in a straight line. Meanwhile, the protein is denatured with detergent (eg sodium dodecyl sulfate, SDS) to make the protein straight and negatively charged.
After the electrophoresis process is completed, a staining process is carried out so that the separated sample molecules can be seen. Ethidium bromide, silver, or "Coomassie blue" (Coomassie blue) colorants can be used for this purpose. If the sample molecule glows in ultraviolet light (for example after it has been "stained" with ethidium bromide), the gel is photographed under ultraviolet light. If the sample molecule contains a radioactive atom, a gel autoradiogram is made.
Molecular Sciences
Molecules called molecular chemistry or molecular physics depend on the focus of research. Molecular chemistry deals with laws governing interactions between molecules, while molecular physics deals with laws that govern the structure and properties of molecules. In practice, the difference is that science is unclear and overlaps.
In molecular science, molecules consist of stable systems consisting of two or more molecules. Polyatomic ions can also sometimes be considered as charged molecules. The term unstable molecule is used to refer to a highly reactive chemical species.
After insertion into the cell, the protein encoded by the DNA fragment can be expressed by the cell in question. Various types of methods can be used to help the expression so that the relevant protein is obtained in large quantities, such as inducible promoters and specific cell-signaling factors. Large amounts of protein can then be extracted from the bacterial cells or eukaryotes.