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Calcium Flux Analysis Protocol for Flow Cytometry - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/calcium.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for calcium flux analysis. Includes: Reagent Preparation; Sample Preparation; Cell Surface Staining; Instrument Set up; Optimizing the Instrument Settings; Recording Data to Apply Compensation; Recording Experimental Data. - [Read Calcium Flux Analysis Protocol for Flow Cytometry]

Imaging of Organelle Membrane Systems and Membrane Traffic in Living Cells - http://www.cshprotocols.org/cgi/content/abstract/2006/29/pdb.prot4603

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Using confocal laser-scanning microscope & GFP fusion proteins in time-lapse imaging to visualize the behavior of organelles and to track membrane-bound transport intermediates that bud off from organelles. Practical issues related to construction & expression of GFP fusion proteins are discussed. Essential for optimizing the brightness and expression levels of GFP fusion proteins so that intracellular membrane-bound structures containing these fusion proteins can be readily visualized. - [Read Imaging of Organelle Membrane Systems and Membrane Traffic in Living Cells]

Ligation of cDNA to Bacteriophage lambda Arms for the Construction of cDNA Libraries - http://molecularstation.com/protocol-links/admin/dir_links_edit.php?action=N

Category: DNA Protocols: cDNA Library Protocols

The final stage of cDNA library construction involves optimizing ligation of the size-fractionated cDNA to bacteriophage {lambda} arms, followed by packaging and plating of the recombinant bacteriophages. - [Read Ligation of cDNA to Bacteriophage lambda Arms for the Construction of cDNA Libraries]

Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/1/pdb.prot4449

Category: Transfection Protocols

Protocol describes transfection of plasmid DNA into mammalian cell lines using electroporation, a process whereby external application of electric pulses induce cell membrane permeability. Cells in suspension and small volume cells are difficult to transfect, whereas adherent cells and large volume cells are relatively easy. Regardless of cell size or phenotype, transfection efficiency increases with a high concentration of cells in a small volume. - [Read Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol]

Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/1/pdb.prot4449

Category: Transfection Protocols: Electroporation Sonoporation Protocols

Protocol describes transfection of plasmid DNA into mammaliancell lines using electroporation, a process whereby externalapplication of electric pulses induce cell membrane permeability.A number of factors can affect electrotransfection efficiency.In general, cells in suspension and small volume cells are difficultto transfect, whereas adherent cells and large volume cellsare relatively easy. - [Read Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol]

Optimizing PCR Protocols - http://www.jax.org/imr/optimize_pcr.html

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

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

Optimizing PCR Protocols - http://www.jax.org/imr/optimize_pcr.html

Category: PCR Protocols: PCR Optimization

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

Optimizing Western Blots: Pre-Transfer Invitrogen - http://www.invitrogen.com/content.cfm?pageid=10713

Category: Protein Protocols: Western Blot Protocol: Troubleshooting Western Blots

Optimizing Western Blots: Pre-Transfer Invitrogen - [Read Optimizing Western Blots: Pre-Transfer Invitrogen]

Polymerase Chain Reaction (PCR) Optimization - http://www.uaf.edu/museum/resumes/cook/mollab/pcr.html

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]

Relative RT-PCR: Determining the Linear Range of Amplification and Optimizing the Primers:Competimer - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4109

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

In the first part of this protocol, the linear range of amplification is determined by carrying out 10 identical PCRs in the presence of [{alpha}-32P]dCTP and stopping one reaction after every two cycles. Amplification products are quantified on a denaturing polyacrylamide gel and the results plotted on a graph (counts per minute vs. cycle number). Total RNA is used as an internal control. - [Read Relative RT-PCR: Determining the Linear Range of Amplification and Optimizing the Primers:Competimer]