What can I do to sequence through a hairpin or secondairy structure?

– Use the BrilliantDye dGTP kit for the sequencing reaction. For optimal results it can be mixed with the default BrilliantDye kits to increase read lenghts. – Add DMSO to a final 5% (v/v) concentration to the sequencing reaction. – Incubate the reaction at 96C for 10 minutes before cycle sequencing.

How can I troubleshoot an issue with Sanger Sequencing?

-A control primer (M13) and template (pGEM) are provided with each BrilliantDye kit. These can be run following the BrilliantDye protocol and can help you determine whether failed reactions are caused by poor template quality or sequencing reaction failure.

What is the difference between the different versions of BrilliantDye?

-The BrilliantDye Terminator v1.1 Cycle sequencing kit is designed for specialty applications that require optimal basecalling adjacent to the primer. The BrilliantDye Terminator v3.1. Cycle sequencing kit is the method of choice for longest reads. For sequencing through G- and GT-rich templates, NimaGen offers the dGTP BrilliantDye kits, optimized for challenging sequence templates.