Biochemical technique is the tool or method which is usedby the biochemists in their research, laboratory and daily bases.They gave an idea tounderstand about the structure of gene, gene expression, bimolecules and theirfunctions such as protein, carbohydrates, lipids, nucleic acid and also help todiagnose the diseases.These techniques are mainly perfusion, cellfractionation or centrifugation, homogenization, purification, chromatography, microscopy,crystallography, enzyme or protein assay, PCR, isotropic tracer techniques,spectrophotometry, colorimetry, electrophoresis, ultra centrifugation,radioimmuno assay, ELISA and biochemical tests.BIOCHEMICAL TECHNIQUES THAT USED MOSTLY:Ø MicroscopyCells are too small to seen by naked eye, in consequence,the microscope is an instrument which is used to analyze the cell, itsorganelles, and even its molecules can also be seen under the Microscope.The Microscope is based on the technique which is called as Microscopy,which portrayed the principle on which microscope work.
There are number ofmicroscopic techniques used to magnify the objects which are included in belowtwo:· Optical Or Light Microscopy· Electron Microscopy Ø Colorimetry It is the technique which isbased on colorimeter, can determined the concentration of unknown sample bymeasuring the absorbance of specific wavelength of light.Ø ChromatographyIt is the technique used toseparate small quantities of complex mixtures and its components which aredepends upon solid(or stationary phase) and liquid, gas (or mobile phase).It is of many types few are as follow:· Ion-exchangechromatography· Gel filteration chromatography· Affinitychromatography· High performanceliquid chromatography· Adsorptionchromatography· Column chromatography Ø Electrophoresis Itis the technique based on the movement of electrical charges under theinfluence of electric current through an electrolyte for the separation of proteins,nucleic acid and peptides. Electrophoresis is of many types:· Gel electrophoresis(agarose, PAGE)· Paper electrophoresis· Cellulose acetatemembrane electrophoresis Ø Centrifugation It is the technique of separation of different componentsof a mixture according to their molecular weight, size and density by spinning orrotating sample on high speed that component becomes settled down andsupernatant obtained through centrifugal force.
It is of many types:· Differential centrifugation· Rate zonal densitygradient centrifugation· Isopycnic densitygradient centrifugation· Ultra centrifugation Ø X-ray Crystallography It is the techniquein which 3 dimensional structures of macromolecules used to determine theirfunctional analysis of biomolecules byusing x-ray diffraction. IMPORTANCE & APPLICATIONSOF BIOCHEMICAL TECHNIQUES: § Biochemical techniques had a great importance in thefield of drug discovery and development.§ In hematology, immunology, pathology it play an importantrole as a diagnostic purpose for many diseases.§ For the separation and brief knowledge of macromolecules,many techniques are used.§ By the help of these techniques we are able to get theknowledge at molecular level like genomics and proteomics.§ Researches on gene expression, genetic modification and geneticengineering are possible only due to these biochemical techniques.