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The "elution time" of a solute is the time between the start of the separation (the time at which the solute enters the column) and the time at which the solute elutes. In the same way, the elution volume is the volume of eluent required to cause elution. Under standard conditions for a known mix of solutes in a certain technique, the elution ...
Column chromatography proceeds by a series of steps. The mobile phase or eluent is a solvent or a mixture of solvents used to move the compounds through the column. It is chosen so that the retention factor value of the compound of interest is roughly around 0.2 - 0.3 in order to minimize the time and the amount of eluent to run the chromatography.
Thin-layer chromatography is used to separate components of a plant extract, illustrating the experiment with plant pigments which gave chromatography its name. In chemical analysis, chromatography is a laboratory technique for the separation of a mixture into its components.
Chromatography employs continuous adsorption and desorption on a packed bed of a solid to purify multiple components of a single feed stream. In a laboratory setting, mixture of dissolved materials are typically fed using a solvent into a column packed with an appropriate adsorbent, and due to different affinities for solvent (moving phase ...
Finally, elution is the process of adding an aqueous solution to the column, allowing the hydrophilic nucleic acid to leave the column and return to solution. This step may be improved with salt, pH, time, or heat. Finally, to capture the eluate/eluent, the column is transferred into a clean microtube prior to a last centrifugation step.
Anion exchange chromatography is commonly used to purify proteins, amino acids, sugars/carbohydrates and other acidic substances [3] with a negative charge at higher pH levels. The tightness of the binding between the substance and the resin is based on the strength of the negative charge of the substance.
Using this method, DNA fragments can be recovered from a particular region of agarose or polyacrylamide gels. The gel piece containing the fragment is excised (cut out from the whole gel) and placed in a dialysis bag with buffer.
A high-performance countercurrent chromatography system. Countercurrent chromatography (CCC, also counter-current chromatography) is a form of liquid–liquid chromatography that uses a liquid stationary phase that is held in place by inertia of the molecules composing the stationary phase accelerating toward the center of a centrifuge due to centripetal force [1] and is used to separate ...