Capillary electrophoresis Flashcards
Capillary electrophoresis
-is a powerful analytical technique used to separate charged molecules based on their size, charge, and interaction with the background electrolyte.
-It’s widely used in biochemistry and analytical chemistry for applications such as DNA and protein analysis.
What is the difference between gel electrophoresis and capillary electrophoresis?
Gel electrophoresis and capillary electrophoresis are both techniques used to separate biomolecules like DNA, RNA, and proteins, but they differ in their separation medium and overall approach:
-Gel electrophoresis is a well-established, simpler technique, often used for routine analysis or educational purposes.
-Capillary electrophoresis offers higher resolution, automation, and versatility, making it a powerful tool for more complex separations and research applications.
A breakdown of capillary electrophoresis:
- Sample preparation: The sample containing the mixture of molecules to be separated is prepared with a buffer solution.
- Injection: A small amount of the sample is injected into one end of a thin, fused-silica capillary tube.
3.Application of voltage: A high voltage is applied across the capillary, creating an electric field.
- Separation: The charged molecules in the sample migrate through the capillary filled with the electrolyte solution at different rates depending on their properties. Smaller, more highly charged molecules move faster than larger, less charged molecules.
- Detection: As the separated molecules emerge from the other end of the capillary, they are detected by a detector, such as a UV absorbance detector or a mass spectrometer.
- Data analysis: The detector generates a signal that is plotted as a graph of time versus signal intensity (electropherogram). The peaks in the electropherogram correspond to the separated molecules. The analysis of the peak area or height allows for identification and quantification of the components in the mixture.
