Dideoxy chain-termination Sanger sequencing has been used in the past 40 years as a gold standard for gene mutation analysis, de novo sequencing, resequencing, confirmation of next-generation sequencing (NGS), and gap filling for NGS. As a mature technology, Sanger sequencing will continue to play an important role in the DNA sequencing field.
All products provided for the Sanger sequencing workflow, see links below, have proven performance and are offered as valuable alternatives for equivalent products from the sequencer manufacturers.
Template Preparation - Preparation of high-quality sequencing templates and primers for setting up sequencing reactions. PCR templates need to be purified or cleaned up to remove dNTPS, primers and DNA polymerase.
Reaction Setup - Use of one primer and one template to linearly amplify and generate nested fluorescently labeled extension products with a single nucleotide difference.
Clean Up - Clean up of completed sequencing reactions to remove unincorporated dyes and other reaction components from the end-labeled extension products.
Sample Loading & Capillary Electrophoresis - Loading extension products with or without resuspension on CE instruments for detecting fluorescent signals, which are converted into DNA sequence using a base calling software.
Regeneration - As a supplementary workflow step for instrument maintenance, regeneration of the capillary array to remove contaminants accumulated on the array after hundreds of CE runs. Regenerated array regains optimal performance.