Sanger Sequencing White Paper a T C Aa T Factors Affecting Sanger Sequencing Robustness and Data Quality

In order to better understand the robustness of our protocols and to identify failure modes for Sanger Sequencing in the UMGC we carried out a series of experiments testing the effects of DNA template concentration, primer concentration, and sample purity on Sanger sequencing data quality. We determined that the UMGC’s Sanger sequencing processes are robust for a wide range of template and primer concentrations, and that the most likely failure modes for submitted samples center around the presence of contaminants. Finally, we compared the contaminant sensitivity profiles of the UMGC processes to two external vendors, ACGT and Genewiz, and found that the effects of template concentration, primer concentration, and contaminants were highly similar across all three providers. These experiments may help to guide researchers in preparing their samples for Sanger sequencing.