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Nucleic Acid Hybridization

Nucleic acid hybridization is a general method in determining the sequence of homologous DNA, it is also used to differentiate order of genes in a haploid set of chromosomes of a particular organism and the size of limitation fragments that contain such sequence. Meaning, it is possible to study the genetic differences between different organisms and individual here on Earth. A nucleic Acid Hybridization analysis uses five laboratory skills the Restriction fragment preparation: where a restriction enzyme is added to a DNA sample, Electrophoresis: separate the restriction fragment from DNA sample, Blotting: single strands are transferred onto special paper or nylon membranes through capillary action, Radioactive probe: it is added to DNA bands, probe is a single-stranded DNA that is complementary to it, Autoradiography: after the probe hybridizes and excess probe is rinsed off, a sheet of photographic film is laid over the gel, radioactivity in the bound probe exposes the film to form an image corresponding to specific DNA bands-the bands containing DNA that base pairs with the probe. The key on detecting specific nucleic sequence in the mixture of different DNA molecules is in the base pairing between complementary strands. Cytosine and guanine is paired by three hydrogen bonds, thymine and adenine is paired by two hydrogen bonds when DNA is heated the DNA attain heat that drive them to vigorous jiggling as the temperature extend to 95° that may lead hydrogen bond to break. The complementary DNA strands separate in a process called denaturation. Labelled probe is added to the denatured DNA, the probe has a sequence that is complementary to the DNA molecule of interest. When the temperature is lower than 65° complementary strand can renaturate by pairing each other. The label of the probe allows the detection of the DNA interest.…...

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