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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…show more content…
Plasmid C has some similarity from plasmid A but the similar sequences are found on the different size in the fragments. The autoradiograph gives us more information than the agarose gel or its own. In Vitro Mutagenesis In vitro mutagenesis a technique used to introduce specific changes into the sequence of a cloned gene. Mutations alter function of protein product. Mutated gene turned to the host cell, is possible to determine function of the missing normal protein by examining what changes occur in the cell. Methods of In Vitro Mutagenesis: 1. After cloning, the double-stranded plasmid is denatured to obtain a single-stranded DNA template that includes the wild-type version of the DNA of interest. 2. A mutagenic primer is allowed to base-pair with the single-stranded template. The primer consists of two regions that are complementary to template regions on either side of a mismatched region – a region where the base sequence of the mutagenic primer is not complementary and will not pair with the base sequence in the template DNA. 3. Addition of DNA polymerase elongates the primer strand to produce a double-stranded plasmid, one strand representing the original genetic information and the other strand representing the new mutant DNA. 4. The vectors (plasmids) are then used to transform bacterial cells, which replicate both strands of the plasmid. When such a transformed cell first divides,
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