DNA Finge - wells will migrate through the gelatin toward...

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DNA Finge rprinting (RFLP Analysis) In RFLP analysis, the DNA of an organism is cut up into fragments using restriction enzymes. A large number of short fragments of DNA will be produced. Restriction enzymes always cut at the same base sequence. Because no two individuals have identical DNA, no two individuals will have the same length fragments. For example, the enzyme EcoRI always cuts DNA at the sequence GAATTC. Different people are going to have different numbers of this particular sequence and will therefore have different fragment lengths. In addition, some of them will be at different locations on the chromosome. Electrophoresis is a technique used to separate the DNA fragments according to their size. They are placed on a sheet of gelatin and an electric current is applied to the sheet. DNA is charged and will move in an electric field toward the positive pole. In the diagram below, holes (wells) in the gelatin can be seen. DNA samples placed in these
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Unformatted text preview: wells will migrate through the gelatin toward the + side after an electric current is applied. The smallest fragments will move the fastest because they are able to move through the pores in the gelatin faster. Bands will be produced on the gelatin where the fragments accumulate. The shortest fragments will accumulate near one end of the gelatin and the longer, slower-moving ones will remain near the other end. In the diagram below, four samples of DNA were placed on the gelatin. After an electric current was applied for a period of time, the fragments separated. Notice that sample D on the right does not match the other three samples. The DNA bands must be stained to make them visible. Ethidium bromide-stained DNA will fluoresce when illuminated with UV light. PCR techniques are used to produce sufficient quantities of DNA for this technique....
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This note was uploaded on 12/15/2011 for the course BIO BSC1010 taught by Professor Gwenhauner during the Fall '10 term at Broward College.

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DNA Finge - wells will migrate through the gelatin toward...

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