Failed Reaction

This peak pattern is the software's attempt at base calling background noise. The signal intensity is very low. Notice the presence of a blue dye blob. This indicates that BigDye was added and the blob peak would be obscured in a normal sequence. A reaction can fail for several reasons: a missing or very dilute reactant, primer binding site mismatches, a Tm of less than 50°C, primer dimers, a binding site inside a large secondary structure fold, or the binding site not actually being present in the construct. Cancel out some of these reasons per your experiment.