12
Electrophoresis
Martha L. Booz
Chemical Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Is the Safest Approach to Working with Acrylamide? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Are the Symptoms of Acrylamide Poisoning? . . . . . . What Is the Medical Response to Accidental Acrylamide Exposure? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . How Can You Dispose of Excess, Unusable Acrylamide? . . What Is the Shelf Life of Acrylamide and Acrylamide …show more content…
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . How Do You Choose a Detergent for IEF or Native PAGE? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Other Additives Can Be Used to Enhance Protein Solubility? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Agarose Electrophoresis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Is Agarose? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Is Electroendosmosis (-Mr or EEO)? . . . . . . . . . . . . . . Are Double-Stranded Markers Appropriate for Sizing Large Single-Stranded (Not Oligonucleotide) DNA? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Causes Nucleic Acids to Migrate at Unexpected Migration Rates? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Causes Commercial Preparations of Nucleic Acid Markers to Smear? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Causes Fuzzy Bands? . . . . . . . . . . . . . . . . . . . . . . . . . . . Elution of Nucleic Acids and Proteins from Gels . . . . . . . . . . . Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Should You Consider before Selecting a Stain? . . . . . . Will the Choice of Stain Affect a Downstream Application? . …show more content…
The wires connecting the cell to the power supply must be in good condition, not worn or cracked, and the banana plugs and jacks must be in good condition, not corroded or worn. Broken or worn wires can cause rapid changes in resistance, slow electrophoresis or a halt in the run. All cables and connectors must be inspected regularly for breaks and wear. The banana plugs on the ends of the wires should be removed from the power supply at the end of the run by pulling them straight out. Grasp the plug, not the wire. If pulled at an angle, the solder joint attaching the banana plugs to the wires can loosen and cause the loss of the electrical circuit. On the cell core, electrode banana posts with flattened baskets do not make good contact with the banana jack in the cell lid, and should be replaced. The banana jacks (female part) in the cell lid should be inspected regularly to make sure there is no corrosion. Before starting an electrophoresis run, dry any liquid on the horizontal surfaces of the cell, especially near the banana plugs and jacks. Any liquid on the horizontal surfaces of the cell can arc during the run, damaging the cell and stopping the electrophoresis.
POLYACRYLAMIDE (PAGE) GELS—BEFORE SELECTING A GEL: GETTING THE BEST RESULTS FOR YOUR PURPOSE Before choosing which gel to use, it is important to consider several questions, all of which can help you choose the gel that will give you the best results