推荐：PCR从引物设计到反应条件的优化
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这份资料以言简意赅的幻灯形式展示了PCR从引物设计到反应条件的优化应注意的问题，相信对您的实验有指导意义。
极力推荐！
http://www.biotechlab.nwu.edu/pe/
包括以下内容：
Introduction - PCR Primer
Components
Primer Design
Primer Design - I
Primer Design - II
Melting Temperature, Tm
Tm Calculations, Nearest Neighbor Analyses I
Tm Calculations, Nearest Neighbor Analyses II
Quantitating Nucleic Acids
Calculating Oligonucleotide Molarity
Reaction Conditions-Background
Conditions Affecting Fidelity
Reaction Parameters That Affect PCR Efficiency
Conditions Affecting Fidelity
Reaction Set Up
Optimization
Optimization of PCR Reactions - Why?
Biochemical Reminder
Buffer Optimization
PCR Primer Concentrations
Template
PCR Product Copy Number Depends on Target Starting Copy Number
Input Copy Number, Number of Cycles, and Product Mass
Mg++
Mg++ in PCR
Mg++ Optimization
Enzyme
Enzyme Choice
Different PCR Enzymes for Different Applications
Different PCR Enzymes Mean Different PCR Buffers
Optimum Mg++ Ranges Depend Upon Polymerase
Effect of [Mg++] and [Enzyme] on Yield
Other Reaction Parameters and Potential Artifacts
The Plateau Effect
The Plateau Effect - I
Plateau Effect - II
Other Artifacts
Three-way competition in PCR
XX Mispriming Models
Pre-PCR Mispriming
Solution to Artifacts: Hot Starts
Hot Start Procedures To Increase PCR Specificity
Manual Hot Start Theory
Manual Hot Start Method
AmpliWaxTM Hot Start Method
Hot Start Enzymatic
Activation Curve of AmpliTaq Gold
Using a Pre-PCR Hold for Activation
Manual Hot Start Versus AmpliTaqTM Gold
