BIOL 3110 Lecture Notes - Restriction Fragment Length Polymorphism, Southern Blot, Reverse Transcription Polymerase Chain Reaction
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This method of analysis does not rely on the mutation having a phenotype change, some genes can be polymorphic, therefore, this is a much more useful and accurate method. (refer to fig 4. 1 genes ix) Restriction site polymorphisms change the sequence of the code and can possibly create or destroy restriction site. The higher the recombination frequency the greater the distance the allele is from the center of the gene and vice versa. Gene targeting is a useful tool for identifying the probability of phenotype appearing (genetic diseases); it can also be used for isolating genes of interest. The first step is to identify the size of the dna insert. Once the size of the insert is determined the genome of interest needs to be mapped via restriction enzymes. The mapped genome can now provide the location of the gene and the actual gene can then be analyzed by southern blot analysis. The next step is to perform heteroduplex mapping:
