optimization Protocols

Category: Real Time PCR

Affymetrix Real Time PCR information and optimization. Yale U. Keck Foundation. - [Read Affymetrix Real Time PCR information and optimization]

Category: Epigenetics and Methylation Protocols: ChIP Chromatin Immunoprecipitation Protocols

Chromatin Immunoprecipitation (ChIP) Assay protocol. Millipore. Includes protocols on Optimization of DNA Shearing, Sonication and PCR for ChIP. - [Read Chromatin Immunoprecipitation (ChIP) Assay]

Category: Transfection Protocols

Protocol should be viewed as a starting point for systematic optimization of transfection mediated by lipofecting agents. Once a positive signal has been obtained from a transfected plasmid carrying a standard reporter gene, optimal conditions for transfection can be established by systematic variation of parameters such as the initial cell density, the amount and purity of DNA, the media and serum, and the time of exposure of the cells to the cationic-lipid-DNA complex. - [Read DNA Transfection Mediated by Lipofection Protocol]

Category: PCR Protocols: PCR Optimization

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]

Category: Cloning Protocols

Protocol can be used to optimize ligation conditions for difficult to clone (e.g. very large) fragments. The principle is to independently characterize the ligation kinetics of the vector and insert DNA fragments and then to combine them in optimal ratios. - [Read Ligation Optimization Protocol]

Category: Chromatography Protocols: Dye Ligand Affinity Chromatography Protocols

Protocol for the optimization of absorption condition for dye-ligand affinity chromotography. Generally, low pH and low ionic strength, absence of phosphate ions, and the presence of divalent metals ions increase the binding of proteins to immobilized triazine dyes. - [Read Optimization of Adsorption Conditions for Dye-Ligand Affinity Chromatography Protocol]

Category: Chromatography Protocols: Dye Ligand Affinity Chromatography Protocols

Protocol for the optimization of elution condition for dye-ligand affinity chromotography. Elution methods used in dye-ligand affinity chromatography may be either selective or nonselective in nature.  Usually, selective elution methods are applied in combination with group-specific adsorbents, such as dye-ligand adsorbents, and nonselective elution methods are used in combination with highly specific adsorbents. - [Read Optimization of Elution Conditions for Dye-Ligand Affinity Chromatography Protocol]

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol for the optimization of imidazole concentrations for immobilized metal-ion affinity chromatography. Most samples from which histidine-tagged proteins are to be purified also contain endogenous protein contaminants that bind to the immobilized metal-ion affinity chromatography (IMAC) adsorbent.  Usually, these proteins bind more weakly than the histidine-tagged protein. - [Read Optimization of Imidazole Concentrations for Immobilized Metal-Ion Affinity Chromatography Protocol]

Category: Cell Biology and Cell Culture: Cell Electroporation Protocols

Optimization of Electroporation Transfection of DNA Protocol into Cell Lines. Eppendorf - [Read Optimization Protocol – Cell Line Electroporation PDF]

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols: PCR Optimization Protocols

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

Category: PCR Protocols: PCR Optimization

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

Category: PCR Protocols: PCR Optimization

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.]

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols: PCR Optimization Protocols

Optimizing PCR Reaction Conditions. Optimizing PCR Tips and Guide. University of Alaska Museum - [Read Polymerase Chain Reaction (PCR) Optimization]

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Histidine-tagged proteins can be purified on prepacked 1-ml immobilized metal-ion affinity chromatography (IMAC) columns without optimization of the separation conditions.  The method allows fast capture of the target protein, although with a lower purity than can be obtained under optimized conditions. - [Read Purification of Histidine-Tagged Proteins Using IMAC Without Parameter Optimization Protocol]

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

Protocol optimized for Cos-1 cells. Successful transfection of each cell type requires optimization of the basic protocol. LIPOFECTAMINE method of transient transfection. Upstate. - [Read Transient Transfection of Cos-1 Cells]