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It is used to separate molecules by some properties
Electrophoresis is the motion of particles relative to some fluid influenced by an electric field. The voltage used will affect this electric field, and in turn affect the movement of particles.
Capillary electrophoresis is a technique used in laboratories to separate molecules based on their charge in order to study and analyze them. Capillary electrophoresis uses an electric charge to force the movement of molecules since each molecule will go a varying distance based on the weight of the molecule and their charge. Some areas of study that use capillary electrophoresis include DNA sequencing and pharmaceutical analysis.
1. WHAT IS ELECTROPHORESIS AND WHAT ARE THE IMPORTANTAPPLICATIONS OF ELECTROPHORESIS?Ans. Movement of charged particle in the electric field either towards cathode or anode whensubjected to an electric current is called electrophoresis.The following factors influence the movement of particles during the electrophoresis.(a) Electric current.(b) Net charge of the particle.(c) Size and shape of the particle.(d) Type of supporting media.(e) Buffer solution.Important Applications of ElectrophoresisThe technique of electrophoresis is used to separate and identify the(i) Serum proteins(ii) Serum lipoproteins(iii) Blood hemoglobins2. WHAT ARE THE DIFFERENT TYPES OF ELECTROPHORESIS?Ans. (a) Moving boundary electrophoresis: This technique was first introduced by TISELIUS in 1937(b) Zone electrophoresis: In this type of electrophoresis different types of supporting mediaare used. These are;(a) Paper electrophoresis(i) Whatman filter paper(ii) Cellulose acetate(b) Gel electrophoresis(i) Agarose.(ii) Polyacrylamide gel (used for the separation of isoenzymes).(iii) SDS-PAGE.(iv) Iso-electric focussing (proteins seperated in a medium possessing a stable pH gradient).(v) Immuno electrophoresis (for the separation of immunoglobulins).
Gel electrophoresis is commonly used in molecular biology for various applications such as DNA fingerprinting, analyzing gene expression, and studying genetic mutations. It is also used in the separation and analysis of proteins, RNA, and DNA fragments in research and diagnostic laboratories.
Some factors affecting electrophoretic mobility include the charge and size of the molecule, the pH and conductivity of the buffer solution, and the strength of the electric field applied. Factors influencing resolution include the gel matrix composition, the voltage and duration of the electrophoresis run, and the type of staining or detection method used. Adjusting these parameters can help optimize separation and resolution of molecules during electrophoresis.
used to separate macromolecules, either nucleic acids or proteins, on the basis of size, electric charge, and other physical properties. Separating strands of DNA
To separate and analyze DNA fragments and protein fragments by weight. If you have digested some bacterial DNA, for instance, then you can tell by running the fragmented DNA in the gel whether you have digested the correct base length.
Agarose gel electrophoresis is used to separate DNA fragments based on size. When an electric current is applied to the gel, DNA molecules move through the pores of the gel at different rates depending on their size, allowing for visualization and analysis of DNA fragments in a sample.
There are many similarities and differences between protein and DNA electrophoresis.Similarities:PAGE protein and DNA electrophoresis both cause separation by size, creating bands that are viewed by the scientist or a machine. The smallest segments more the fastest due to less friction with the surface of their medium or equipment.The movement of charges through the medium is what causes the DNA or proteins to move. Electrons move from the negative to positive end of the gel or capillary tube.Differences:In PAGE protein electrophoresis, a polyacrylamide gel is used to prevent convection from altering the movement of the proteins. If the proteins are charged, and there is a worry that the charge will affect the mobility of the protein segments, 1% SDS can be added to get rid of the mass/charge issue. This way, only the mass of the segment determines how far it moves. In DNA capillary electrophoresis, the size of the capillary is so small that it does not have room for convection to occur (it is only 20-50 microns wide). Thus, there is no medium in the capillary but DNA itself.In protein electrophoresis, the proteins are often dyed so their movement can be viewed with the naked eye, or a machine. With DNA capillary electrophoresis, DNA strands are made through DNA replication with dNTPs that are fluorescently labeled for the different nucleotides. Each base is labeled a different color. A fine laser lights up the DNA strand in the capillary tube and reads what color fluoresces. This is how the nucleotide is identified.Protein PAGE electrophoresis is used to determine the purity of a protein sample. It can also be used to see how large the chains are that make up a multi-chain protein if a denaturing agent is added. DNA electrophoresis is used to get the order of nucleotides in a DNA sequence. It is done by chopping the DNA sequence into many smaller bits and sequencing them, then putting them back together by lining them up according to sequence overlaps. This is called the "shotgun" method. Protein electrophoresis can figure out the order of about 15-20 amino acids by a similar method, but DNA electrophoresis can get up to 1000 nucleotides (~300 amino acids). DNA electrophoresis is limited by the low probability that the DNA sequence would be cut into a segment greater than 1000 nucleotides.
The idea is to work with the same variables, but it is possible that some of the variables are missing in some of the equations.
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