PCR Optimization

Calculating Concentrations for PCR. Ed Rybicki. Calculating Primer and Nucleotide Concentrations for PCR - [Read Calculating Concentrations for PCR]

Protocol for enhancing PCR of very difficult regions. - [Read Enhancing PCR of Very Difficult Regions Protocol]

Method for amplifying DNA enzymatically by the polymerase chain reaction (PCR), including procedures to quickly determine conditions for successful amplification of the sequence and primer sets of interest, and to optimize for specificity, sensitivity, and yield.  The first step of PCR simply entails mixing template DNA, two appropriate oligonucleotide primers, Taq or other thermostable DNA polymerases, deoxyribonucleoside triphosphates (dNTPs), and a buffer. - [Read Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization Protocol]

Method describes how to modify the termini of PCR products by introducing restriction sites and other features.  To reduce the chance of contamination with exogenous DNAs, prepare and use a special set of reagents and solutions for PCR only.  Bake all glassware for 6 hours at 150°C and autoclave all plasticware. - [Read Genetic Engineering with PCR Protocol]

Optimizing PCR Protocols. The Jackson Laboratory. - [Read Optimizing PCR Protocols]

Procedure details the establishment of an amplification procedure for GC-rich sequences.  The DNA fragments of interest are amplified in the presence of either 5% DMSO, 1 M betaine, 2 M betaine, 1 M betaine, and 5% DMSO; 2 M betaine and 5% DMSO; 0.4 M tetramethylene sulfone; or without any of the enhancers. - [Read PCR Amplification of Highly GC-Rich Regions Protocol]

A Great PCR and Multiplex Guide. Topics for PCR optimization. Primer conditions, PCR temperatures, etc. Octavian Henegariu. - [Read PCR and Multiplex Guide]

PCR primer design and pcr reaction optimization. Ed Rybicki. Factors Affecting the PCR, Denaturing Temperature and Time, Annealing Temperature and Primer Design, Primer Length, Degenerate Primers, Elongation , Temperature and Time, Reaction Buffer, Cycl - [Read PCR primer design and pcr reaction optimization.]

PCR PRIMER DESIGN AND REACTION OPTIMISATION: Molecular Biology Techniques Manual - Rybicki - [Read PCR PRIMER DESIGN AND REACTION OPTIMISATION: Molecular Biology Techniques Manual]

The requirement of an optimal PCR reaction is to amplify a specific locus without any unspecific by-products. Therefore, annealing needs to take place at a sufficiently high temperature to allow only the perfect DNA-DNA matches to occur in the reaction. P - [Read PCR Program Design]

PCR Tips and Tricks - General Guidelines and Troubleshooting PCR polymerase chain reaction. The Genomic Variation Laboratory, U.C Davis - [Read PCR Tips and Tricks]