Fragment analysis generates a size estimate for DNA fragments relative to a size standard of DNA fragments with known lengths. The size standard is combined with the sample of interest and co-injected on the capillary electrophoresis system.
What is Fragment Analysis
- Labeling fragments with fluorescent dyes. Multiple different colored fluorescent dyes can be detected in one sample. One of the dye colors is used for a labeled size standard present in each sample. The size standard is used to extrapolate the base-pair sizes of the sample product peaks.
- Amplifying the labeled fragments using polymerase chain reaction (PCR) on a thermal cycler.
- Separating the fragments by size using capillary electrophoresis.
- Analyzing the data using software to determine:
- Size : The analysis software uses the size standard in each sample to create a standard curve for each sample. It then determines the relative size of each dye‐ labeled fragment in the sample by comparing fragments with the standard curve for that specific sample.
- Genotype : The analysis software assigns allele calls based on user‐ defined makers (loci).