What controls are used in capillary electrophoresis?
It involves the separation of peptides by application to a fused silica capillary (typically 100 cm×100 μm). Electrophoretic mobility is controlled by an external electric field and selectivity can be manipulated by a number of factors including solvent pH, ionic strength, and other additives.
What factors affect separation in capillary electrophoresis?
Capillary Zone Electrophoresis (CZE) The separation is based on the differences in electrophoretic mobility, which is directed proportional to the charge on the molecule, and inversely proportional to the viscosity of the solvent and radius of the atom.
What is the detector in capillary electrophoresis?
Detection. Separation by capillary electrophoresis can be detected by several detection devices. The majority of commercial systems use UV or UV-Vis absorbance as their primary mode of detection. In these systems, a section of the capillary itself is used as the detection cell.
What causes split peaks in an Electropherogram?
Split peaks are due to the main peak being split into two peaks caused by the Taq polymerase activity that causes the addition of a single “A” to the terminus of the amplified product (“N+1” band).
What is the purpose of capillary electrophoresis?
Capillary electrophoresis (CE) is the primary methodology used for separating and detecting short tandem repeat (STR) alleles in forensic DNA laboratories worldwide. This chapter examines the general principles and components of injection, separation, and detection of STR alleles using CE.
What are the factors that affect electrophoresis?
Factors affecting electrophoresis include characteristics of the ion or molecule itself, the environment (buffer) in which the molecule or ions are being studied, and the applied electrical field. These factors specifically affect the migration rates of molecules in the sample during electrophoresis.
What factors affect MOPH electrophoresis?
2 Factors Affecting Electrophoretic Mobility Charge – The higher the charge, the greater the mobility. Size – The bigger the molecule, the greater the frictional and electrostatic forces exerted on it by the medium, i.e., larger particles have smaller electrophoretic mobility compared to smaller particles.
What is a stutter peak?
Stutter peaks are small peaks that occur immediately before or after a real peak. During the PCR amplification process, the polymerase can lose its place when copying a strand of DNA, usually slipping forwards or backwards four base pairs.
How to troubleshoot DNA sequencing by capillary electrophoresis chemistry?
206 DNA Sequencing by Capillary Electrophoresis Chemistry Guide Chapter 8 Troubleshooting Pull-up caused by overloading the capillaries with too much product. Review DNA quantity. Use standard run modules Click the Annotationtab and examine the Ave Signal Intensity.
What are the symptoms of electrophoresis at Notre Dame?
Electrophoresis issues (likely in multiple lanes and/or runs): Carryover from contaminated septa. Replace septas and change buffer, water, and waste. Electrical noise. Check the uninterruptible power supply (UPS). Contaminated water or buffer because of dirty containers, microbial growth, or use of tap water for cleaning.
How to troubleshoot common amplification errors and issues?
Trouble Shooting Tools Common Capillary Electrophoresis Instrument Issues and Observations Trouble Shooting 3500 Issues Common Amplification Observations 2/8/2013| © Life Technologies™4 Troubleshooting Tools