Fragment Analysis

Fluorescently labeled fragments are detected using the Applied Biosystems 3730xl Genetic Analyzer and then interpreted using the GeneMapper analysis software. Up to five different colored fluorescent dyes can be detected in one sample well. One of the dye colors is used for a labeled size standard that is added to each lane to allow comparison of samples from lane to lane. The analysis software uses the size standard to create a standard curve for each lane and then determines the length of each dye-labeled fragment by comparing it with the standard curve for that specific lane. Accuracy of less than one base difference is seen between sample replicates. Higher throughput can be achieved by multiplexing different dyes and fragment sizes together.