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Introduction to Quantitative PCR - Agilent

Methods and Applications GuideIntroduction to Quantitative PCR Introduction to Quantitative PCR Methods and Applications Guide IN 70200 D US and Canada Orders: 800-227-9770 x3 Technical Service: 800-227-9770 x2 For a list of worldwide distributors, please visit Agilent Technologies, Inc. 2012 Table of Contents Table of Contents .. 2 Introduction to Quantitative PCR .. 1 Real-Time vs. Endpoint Quantitative PCR .. 2 Experimental Design .. 5 Methods of Quantification .. 6 Standard Curve .. 6 Relative Quantification .. 8 QPCR Chemistry Options .. 9 Primer and Probe Design .. 16 Dye and Quencher Choice .. 20 QPCR Reagent Choice .. 21 Probe and Primer Synthesis .. 24 Reference Dye Considerations .. 24 Nucleic Acid Sample Preparation .. 26 DNA .. 26 RNA .. 26 Total RNA vs. mRNA .. 26 Measuring RNA Quality .. 27 Storage of RNA .. 29 Using Cell Lysates in Real-Time QPCR .. 30 Reverse Transcription Considerations.

Introduction to Quantitative PCR Whether you are a novice or experienced user, our goal is to ensure that you are running quantitative PCR (QPCR) experiments

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