Optimizing the Polymerase Chain Reaction
In this advanced PCR protocol, students work in teams to optimize one of two components in a PCR reaction: magnesium chloride or primer concentration. Students set up 7 reactions per team, each containing a different concentration of magnesium chloride or primer. Before starting the
procedures, students must first calculate the necessary volumes of each component to be added into each reaction. This exercise is very math oriented and is an excellent way to demonstrate the prevalent use of mathematics in biotechnology. After PCR, electrophoresis, staining, and photographing, students analyze their gels to determine the optimum magnesium chloride and primer concentration for the PCR amplification of a segment of the acetylcholinesterase gene from cow DNA.
PCR Optimization Student Laboratory Guide
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