Transcription of PCR Optimization Student Guide 2012 - Babec
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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 ..29 Interpretation of Results Role of Magnesium ..30 Role of Primers ..32 Acknowledgements ..33 !PCR Optimization Student Guide Fall 2012 1 !Optimizing the Polymerase Chain Reaction Frank H. Stephenson, Maria C. Abilock Applied Biosystems Babec Introduction The Polymerase Chain Reaction (PCR) is a powerful technique used for the amplification of a specific segment of a nucleic acid.
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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