Sanger Sequencing

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.

The experimental workflow for Sanger sequencing includes:

Sanger Sequencing Products

SupreDye™ Cycle Sequencing Kits

High-performance alternative for BigDye™ Terminator v. 3.1 or 1.1 Cycling Sequencing Kit

ADS™ BD-XT Purification Kit

For efficient contaminant removal

TruPure™ Formamide

For high-performance sequencing sample resuspension

PwrPOP™ Polymers

For efficient contaminant removal

ADS™ Capillary Regeneration Kit

For sustaining high-quality sequencing

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

Cleanup of completed sequencing reactions to remove unincorporated dyes and other reaction components from the end-labeled extension products.

Sample Loading & CE

Loading extension products with or without resuspension on CE instruments for detecting fluorescent signals, which are converted into DNA sequence using a basecalling 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.