CHAPTER ONE
1.0 INTRODUCTION
For many years, people died of many diseases they have no knowledge of nor do they have idea of how to curve. It was not clear to them if the death, was as a result of a single disease or multiple of diseases. (1)
People worked tirelessly and laboriously but the produce from their farm was not enough to feed their family not to talk of selling them to generate a sustainable income.
Many couple witnessed sudden death of their child at birth or at their teen, without knowing the cause nor how to avert it.
The advent of the knowledge of electrophoresis threw light on the genotypical constitution of cells. This gave rise to hybrid as a result of cross bred, which eventually solved the problem of low yield (harvest).
The knowledge also goes a long way to diagnose the causes of different diseases that causes death, the likes of cancer called multiple myelona, sickle cell, diseases at birth, chronic liver disease etc.
The knowledge did not only unravel the causes but also led to the development of authentic cure. Also intending couple’s eyes were opened to their genotype status, giving them room to decide on whether to go on with their marriage or not, as the likelihood or otherwise of having sickler as a child is made know to them.
CHAPTER TWO
2.0 LITERATURE REVIEW
Electrophoresis is the migration of charged particles under the influence of an electric field. Electrophoresis is a method that separates macromolecules – either nucleic acids or protein – on the basis of size, electric charge and other physical properties. ELECTRO refers to the energy of electricity. PHORESIS, from the Greek verb PHOROS, means “to carry across”. Thus, electrophoresis refers to the technique in which molecules are forced across a span of gel, motivated by an electric current. In 1955, smithies was the first to introduced a major development in electrophoresis by using remarkable resolving power of starch gel electrophoresis for serum protein.
Electrophoresis machine is a machine that supply a voltage of 200V its tank containing cathode electrode, anode electrode and a buffer solution of pH 8.6 and the blood serum to be tested. Many important biological molecules such as Amino acids, exist at any given pH solution as electrically charged particle either as cations (+) or anion (-). Depending on the nature of the net charge, the charged particles will migrate either to the cathode or to the anode.
To completely understand the separation of charged particles in electrophoresis, it is important to look at some simple equation relating to electrophoresis. When potential difference (voltage) is applied across the electrodes, it generate a potential gradient (E), which is the applied voltage (V) divided by the distance (d) between the electrodes.
E = v/d
When the potential gradient E is applied, the some on a molecule bearing a charge of q coulombs is Eq newtons.
F = Eq.
It is this force that drives a charged molecule towards an electrode.
There is also a frictional resistance that slows down the movement of this charged molecule. The frictional force is a measure of the hydrodynamic size of the molecule, the shape of the molecule, the pore size of the medium in which electrophoresis is taking place and viscosity of the buffer. The velocity (v) of a charged molecule in an electric field is given by the equation.
V = Eq
f
where f is the frictional co-efficient.
In electrophoresis, the force moving the micro molecule (nucleic acids or proteins) is the electrical potential, E. The electrophoresis mobility (???) of an ion is the ratio of the velocity of the particle, (v) to the electrical potential.
= V
E
Electrophoresis mobility is also equal to the net charge of the molecule, Z divided by the frictional coefficient, f.
= Z
f
When a potential difference is applied, molecules with different overall charges will begin to separate due to their different electrophoresis mobilities. Even molecules with similar charges will begin to separate if they have different molecular sizes. Since they will experience different frictional forces. (1)
The current in a solution between the electrodes is conducted mainly by the buffer ions with a small proportion being conducted by the sample ions. Ohm’s law expresses the relationship between current (I), voltage (V), and resistance (R).
R = V
1
This equation demonstrates that it is possible to accelerate an electrophonetic separation by increasing the applied voltage, which would result in a corresponding increase in the current flow. The distance migrated will be proportional to both current and time.
However, increasing the voltage would ignore one of the major problems for most forms of electrophoresis, namely the generation of heat. During electrophoresis the power (W, watts) generated in the supporting medium is given by:
W = 12 R
APPLICATIONS & USE OF ELECTROPHORESIS
As mentioned earlier the use of Electrophoresis and electrophoresis machine is countless as it is in wide use in pharmaceutical industries, clinical laboratories, science and Genetic research laboratories, clinical research laboratories, genetical finger printing laboratories etc.
The knowledge of electrophoresis have been use extensively in the identification of the genotypical constitution of human, in the identification of particular DNA molecules by the band pattern they yield in gel electrophoresis, an individual distinctive as infinger print, screening for a rare type of cancer called multiple myelona, to diagnose abnormal protein level cause by chronic liver disease, to screen for sickle cell disease at birth, to bring about hybrid species. Another use is the isolation and purification of individual fragments containing interesting genes, to determine the genetic difference and evolutionary relationship among species of plant and animals.
