Terminal Restriction Fragment Length Polymorphism (T-RFLP) Analysis by Capillary Electrophoresis (CAT#: STEM-ET-0306-ZJF)

Capillary Electrophoresis (CE) is physical method of analysis which performs in a separation channel of elastic quartz capillary, under the influence of a high voltage direct current field. Charged analytes dissolved in an electrolyte solution are separated based on differences in mobility and/or distribution behavior of components. The migration velocity of an analyte under an electric field is determined by the electrophoretic mobility of the analyte and the electro-osmotic mobility of the buffer inside the capillary. The electrophoretic mobility of a solute depends on the characteristics of the solute (electric charge, molecular size and shape) and those of the buffer in which the migration takes place (type and ionic strength of the electrolyte, pH, viscosity and additives). Capillary electrophoresis provides greater resolution, higher sensitivity and online detection. It enables single-cell analysis and even single-molecule analysis, optimizing separation and analysis of biological macromolecules.

Genomics, T-RFLP

1. Genomic DNA extraction
2. Fluorescent PCR amplification
3. Enzyme digestion performed by restriction endonuclease
4. Capillary electrophoresis detection
5. Raw data analysis

• Capillary electrophoresis apparatus
• Sample solution
• Buffer solution

Sample requirements:
PCR products with fluorescent labeling:> 30ul, transported by ice pack.
Genomic DNA: Concentration > 20-30ng/ul, OD260/280 about 1.7-2.1, transported by ice pack.
Soil: Dry or wet, dry soil effect is better, quality > 5g