Heating and Mixing
Electrophoresis and Blotting
Polyacrylamide Gel Electrophoresis
Agarose Gel Electrophoresis
PCR & qPCR Thermal Cycler
Thermal Cycler (PCR)
Real-time Thermal Cycler (qPCR)
PCR Workstations & Cabinets
UVP BioImaging Systems
UVP Benchtop Transilluminators
Electrophoresis & Blotting
Shaker & Mixer
Orbital Shaking Incubator
Water Purification System
Aermax - Air Purification
Medical Oxygen Concetrators
-150°C Cryogenic Freezer
-86°C Ultra Low Temp Freezer
-40°C Low Temp Freezer
-18 ~ -25°C Biomedical Freezer
-20°C Biomedical Freezer
4° ± 1°C Blood Bank Refrigerators
2~8°C Pharma Refrigerators
2~8°C ICE Lined Refrigerators
-25°C ~ + 4°C Mobile Freezer/Collers
20~24°C Blood Platelet Incubators
Electrophoresis is carried out under constant voltage or current or power. The migration rate should be kept constant through a constant power supply. If the increased current is supplied, there is increased heat production through resistance, which causes thermal agitation of dissolved ions. There will be increased evaporation of water from the apparatus. This will lead to an increase in the concentration of ions in the buffer.
Buffer establishes the electrical charge on the solute and pH of the system. It is used in carrying the electrical current.
The ideal buffer:
Citrate, acetate, formate, and phosphate are acidic buffers used for lower pH. Tris, tricine, and borate are basic buffers used for maintaining high pH. The buffers which are used are made up of monovalent ions as the valency (ionic strength) and molality is equal. The prepared buffers can serve as a good platform for the growth of microorganisms, so they should be properly refrigerated when they are not used. Cold buffer can be used in the process as it provides increased resolution of samples and decreased evaporation of the solvent in the process. Large volumes of the buffer can be reused up to four times, but the smaller volumes can be discarded immediately. The higher ionic strength of the buffer is useful in producing a sharper resolution of samples, though there is a risk of damage to heat-labile compounds as heat is generated much.
Slabs, sheets, columns of starch, polyacrylamide, agarose, the membrane of cellulose acetate are used as supporting media. It is in a colloid form, made up of more than 90% of water. It acts as a molecular sieve, through which the separation of molecules is done. It is porous, allowing small molecules to pass through while obstructing the movement of larger molecules. It should be electrically neutral. Nowadays, agarose gel is mainly used as a support medium to carry out electrophoresis.
Separates biomolecules on the basis of molecular size and surface charge
It is not used now as preparation of gel showing reproducibility is pretty difficult
The separation of molecules is based only on mass to charge ratio as all proteins can pass through the pore of the gel. It is capable of separating proteins into five zones only. There is a lower affinity for proteins, which provides good clarity after drying. Endosmosis is negligible and it is devoid of ionizable groups, being neutral. The downside is the recovery of DNA is affected by inhibitors.
It is a plastic box or tank, which is filled with a buffer for the migration of biomolecules. It has a transparent cover, which facilitates easy viewing of the migration process. It is connected to a power source.
The trays and containers are used for the purposes of staining and destaining gels. Both open form and lidded boxes are available. Generally, they have a polypropylene base. They are tightly fitted and clear. They are resistant to stains and solvents.
There are two platinum electrodes, which facilitate the separation of molecules by attracting opposite charges. An anode is used for binding to positive ions and a cathode binds negative ions.
The gel is poured after dissolving in the solvent into a gel caster, which holds the gel and is kept inside the apparatus. A comb is used to put the wells for the loading of samples.
Each and every part of the electrophoretic apparatus is useful and significant in one or another way. Depending on the type of electrophoresis, the reagents used and parts may vary. Concentration may be varied depending on the status of the sample. The methodology for the technique is given, yet it differs slightly for different types of electrophoresis. While doing the experiment, following optimized protocol is recommended.