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Autoradiography
Autoradiography is a modification of the radioactive tracer technique. In this technique, the radioactivity is detected by a thin film of photographic emulsion containing silver compounds which is put over a labelled cell or tissue. The rays emitted by radioisotope from the cell or tissue expose the photographic emulsion like a photographic film which is developed in a dark room. Silver grains are seen as black dots on parts of the specimen where radioisotope is accumulated.
Autoradiography reveals the metabolic processes of the cell. For example, by labelling cells with 3H thymidine, it is found that silver grains are localised in nucleus, which shows that DNA is synthesised in the nucleus. Similarly, by labelling cells with 3H uridine, silver grains are first seen in nucleus and then after some time these grains appear in the cytoplasm. This clearly demonstrates that RNA is initially synthesised in the nucleus but later on migrates to the cytoplasm.Different radioisotopes are used for localising different macromolecules. As stated earlier, cells are labelled with 3H thymidine for the studX of DNA metabolism, 3H uridine to study RNA metabolism, tritiated amino acids for examining the synthesis of proteins, treated monosaccharides such as 3H-mannose and 3H-fucose for examining the synthesis of polysaccharides.
Clonig,plasmid vectors,lambda&M13based vectors
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