Cross Site Evaluation of Sanger Sequencing Dye Chemistries.

: Sanger sequencing remains an essential tool utilized by researchers and despite competition from commercial providers, many sequencing core facilities continue to offer Sanger sequencing services to their customer base. By reducing costs and providing rapid turnaround times, in-house Sanger sequencing remains a viable core service, often helping to subsidize more costly services such as next generation sequencing. While Applied Biosystems™ BigDye Terminator chemistry was once the only solution available for Sanger DNA sequencing, several new products employing novel dye chemistries and reaction configurations have entered the market. Currently, it is unclear how these new chemistries perform on various DNA templates, including difficult templates or their amenability to commonly employed cost-saving measures such as dye dilution and reaction miniaturization. With this goal in mind, we compared the quality of Sanger sequencing data produced by kits available from several vendors using control and difficult-to-sequence DNA templates under various reaction conditions. This study will serve as a valuable resource to core facilities conducting Sanger sequencing, providing guidelines on appropriate protocols to use with each kit and determining the most cost effective solutions for Sanger sequencing while maintaining high quality results.