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PCR Optimization Student Guide 2012 - Babec

PCR Optimization Table of Contents Fall 2012 Optimizing the Polymerase Chain Reaction Introduction ..1 Review of Mathematics ..3 Solving Problems of Dilution and Concentration: Two Approaches ..4 Experiment Overview ..7 Calculations Part 1: Overview of PCR Reaction Components ..8 Part 2: Varying the Magnesium Chloride Concentration ..10 Part 3: Varying the Primer Concentration ..15 Laboratory Important Laboratory Practices ..20 Varying Magnesium Concentration ..21 Varying Primer Concentration ..24 Agarose Gel Electrophoresis ..27 Electrophoresis of Amplified DNA ..28 Staining and Photographing Agarose Gels.

PCR Optimization Student Guide Fall 2012 2 ! Figure 1. An optimized PCR amplification will produce a single, bright band on a gel (as seen in the left-most lane).

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