Optimizing the Polymerase Chain Reaction

Inpcr 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 protocol, 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 optimal magnesium chloride and primer concentration for the PCR amplification of a segment of the acetylcholinesterase gene from cow DNA.

Advanced: This lesson requires math skills and proficient to advanced pipetting skills. It is appropriate for advanced biology or biotechnology elective classes after a successful completion of the Introduction to PCR lesson.


PCR Optimization Student Laboratory Guide

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